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Recent Articles in PLoS Medicine / Public Libary of Science

Costa DB, Halmos B, Kumar A, Schumer ST, Huberman MS, Boggon TJ, Tenen DG, Kobayashi S
BIM mediates EGFR tyrosine kinase inhibitor-induced apoptosis in lung cancers with oncogenic EGFR mutations.
PLoS Med. 2007 Oct 30;4(10):1669-79; discussion 1680.
BACKGROUND: Epidermal growth factor receptor (EGFR) mutations are present in the majority of patients with non-small cell lung cancer (NSCLC) responsive to the EGFR tyrosine kinase inhibitors (TKIs) gefitinib or erlotinib. These EGFR-dependent tumors eventually become TKI resistant, and the common secondary T790M mutation accounts for half the tumors with acquired resistance to gefitinib. However, the key proapoptotic proteins involved in TKI-induced cell death and other secondary mutations involved in resistance remain unclear. The objective of this study was to identify the mechanism of EGFR TKI-induced apoptosis and secondary resistant mutations that affect this process. METHODS AND FINDINGS: To study TKI-induced cell death and mechanisms of resistance, we used lung cancer cell lines (with or without EGFR mutations), Ba/F3 cells stably transfected with EGFR mutation constructs, and tumor samples from a gefitinib-resistant patient. Here we show that up-regulation of the BH3-only polypeptide BIM (also known as BCL2-like 11) correlated with gefitinib-induced apoptosis in gefitinib-sensitive EGFR-mutant lung cancer cells. The T790M mutation blocked gefitinib-induced up-regulation of BIM and apoptosis. This blockade was overcome by the irreversible TKI CL-387,785. Knockdown of BIM by small interfering RNA was able to attenuate apoptosis induced by EGFR TKIs. Furthermore, from a gefitinib-resistant patient carrying the activating L858R mutation, we identified a novel secondary resistant mutation, L747S in cis to the activating mutation, which attenuated the up-regulation of BIM and reduced apoptosis. CONCLUSIONS: Our results provide evidence that BIM is involved in TKI-induced apoptosis in sensitive EGFR-mutant cells and that both attenuation of the up-regulation of BIM and resistance to gefitinib-induced apoptosis are seen in models that contain the common EGFR T790M and the novel L747S secondary resistance mutations. These findings also suggest that induction of BIM may have a role in the treatment of TKI-resistant tumors. [Abstract/Link to Full Text]

Matthews FE, Stephan BC, Bond J, McKeith I, Brayne C
Operationalization of mild cognitive impairment: a graphical approach.
PLoS Med. 2007 Oct 30;4(10):1615-9. [Abstract/Link to Full Text]

Williamson J, Proctor C
Should the health community promote smokeless tobacco (snus): comments from British American Tobacco.
PLoS Med. 2007 Oct 30;4(10):1703-4; author reply 1704-5. [Abstract/Link to Full Text]

Chapman S
Should the Health Community Promote Smokeless Tobacco (Snus): Author's Reply.
PLoS Med. 2007 Oct 30;4(10):e299. [Abstract/Link to Full Text]

Szalavitz M
Tobacco substitutes: snus and harm reduction.
PLoS Med. 2007 Oct 30;4(10):1703. [Abstract/Link to Full Text]

Silverman J, Decker M
The US anti-prostitution pledge: a call for cooperation.
PLoS Med. 2007 Oct 30;4(10):1702; author reply 1702-3. [Abstract/Link to Full Text]

Piot P, Greener R, Russell S
Squaring the circle: AIDS, poverty, and human development.
PLoS Med. 2007 Oct 23;4(10):1571-5. [Abstract/Link to Full Text]

Thirty ways to improve the health of the world's poorest people.
PLoS Med. 2007 Oct 23;4(10):1555-6. [Abstract/Link to Full Text]

Obrist B, Iteba N, Lengeler C, Makemba A, Mshana C, Nathan R, Alba S, Dillip A, Hetzel MW, Mayumana I, Schulze A, Mshinda H
Access to health care in contexts of livelihood insecurity: a framework for analysis and action.
PLoS Med. 2007 Oct 23;4(10):1584-8. [Abstract/Link to Full Text]

Pemberton S, Gordon D, Nandy S, Pantazis C, Townsend P
Child rights and child poverty: can the international framework of children's rights be used to improve child survival rates?
PLoS Med. 2007 Oct 23;4(10):1567-70. [Abstract/Link to Full Text]

Bennett GG, McNeill LH, Wolin KY, Duncan DT, Puleo E, Emmons KM
Safe to walk? Neighborhood safety and physical activity among public housing residents.
PLoS Med. 2007 Oct 23;4(10):1599-606; discussion 1607.
BACKGROUND: Despite its health benefits, physical inactivity is pervasive, particularly among those living in lower-income urban communities. In such settings, neighborhood safety may impact willingness to be regularly physically active. We examined the association of perceived neighborhood safety with pedometer-determined physical activity and physical activity self-efficacy. METHODS AND FINDINGS: Participants were 1,180 predominantly racial/ethnic minority adults recruited from 12 urban low-income housing complexes in metropolitan Boston. Participants completed a 5-d pedometer data-collection protocol and self-reported their perceptions of neighborhood safety and self-efficacy (i.e., confidence in the ability to be physically active). Gender-stratified bivariate and multivariable random effects models were estimated to account for within-site clustering. Most participants reported feeling safe during the day, while just over one-third (36%) felt safe at night. We found no association between daytime safety reports and physical activity among both men and women. There was also no association between night-time safety reports and physical activity among men (p = 0.23) but women who reported feeling unsafe (versus safe) at night showed significantly fewer steps per day (4,302 versus 5,178, p = 0.01). Perceiving one's neighborhood as unsafe during the day was associated with significantly lower odds of having high physical activity self-efficacy among both men (OR 0.40, p = 0.01) and women (OR 0.68, p = 0.02). CONCLUSIONS: Residing in a neighborhood that is perceived to be unsafe at night is a barrier to regular physical activity among individuals, especially women, living in urban low-income housing. Feeling unsafe may also diminish confidence in the ability to be more physically active. Both of these factors may limit the effectiveness of physical activity promotion strategies delivered in similar settings. [Abstract/Link to Full Text]

Poma SR, Soto MV, Chahua Jde L, Poma MR
Which single intervention would do the most to improve the health of those living on less than $1 per day? Interview by Gavin Yamey.
PLoS Med. 2007 Oct 23;4(10):1557-60. [Abstract/Link to Full Text]

Rollins N
Food insecurity--a risk factor for HIV infection.
PLoS Med. 2007 Oct 23;4(10):1576-7. [Abstract/Link to Full Text]

Weiser SD, Leiter K, Bangsberg DR, Butler LM, Percy-de Korte F, Hlanze Z, Phaladze N, Iacopino V, Heisler M
Food insufficiency is associated with high-risk sexual behavior among women in Botswana and Swaziland.
PLoS Med. 2007 Oct 23;4(10):1589-97; discussion 1598.
BACKGROUND: Both food insufficiency and HIV infection are major public health problems in sub-Saharan Africa, yet the impact of food insufficiency on HIV risk behavior has not been systematically investigated. We tested the hypothesis that food insufficiency is associated with HIV transmission behavior. METHODS AND FINDINGS: We studied the association between food insufficiency (not having enough food to eat over the previous 12 months) and inconsistent condom use, sex exchange, and other measures of risky sex in a cross-sectional population-based study of 1,255 adults in Botswana and 796 adults in Swaziland using a stratified two-stage probability design. Associations were examined using multivariable logistic regression analyses, clustered by country and stratified by gender. Food insufficiency was reported by 32% of women and 22% of men over the previous 12 months. Among 1,050 women in both countries, after controlling for respondent characteristics including income and education, HIV knowledge, and alcohol use, food insufficiency was associated with inconsistent condom use with a nonprimary partner (adjusted odds ratio [AOR] 1.73, 95% confidence interval [CI] 1.27-2.36), sex exchange (AOR 1.84, 95% CI 1.74-1.93), intergenerational sexual relationships (AOR 1.46, 95% CI 1.03-2.08), and lack of control in sexual relationships (AOR 1.68, 95% CI 1.24-2.28). Associations between food insufficiency and risky sex were much attenuated among men. CONCLUSIONS: Food insufficiency is an important risk factor for increased sexual risk-taking among women in Botswana and Swaziland. Targeted food assistance and income generation programs in conjunction with efforts to enhance women's legal and social rights may play an important role in decreasing HIV transmission risk for women. [Abstract/Link to Full Text]

Unger A, Riley LW
Slum health: from understanding to action.
PLoS Med. 2007 Oct 23;4(10):1561-6. [Abstract/Link to Full Text]

Avilés W, Ortega O, Kuan G, Coloma J, Harris E
Integration of information technologies in clinical studies in Nicaragua.
PLoS Med. 2007 Oct 23;4(10):1578-83. [Abstract/Link to Full Text]

Rosen S, Fox MP, Gill CJ
Patient retention in antiretroviral therapy programs in sub-Saharan Africa: a systematic review.
PLoS Med. 2007 Oct 16;4(10):e298.
BACKGROUND: Long-term retention of patients in Africa's rapidly expanding antiretroviral therapy (ART) programs for HIV/AIDS is essential for these programs' success but has received relatively little attention. In this paper we present a systematic review of patient retention in ART programs in sub-Saharan Africa. METHODS AND FINDINGS: We searched Medline, other literature databases, conference abstracts, publications archives, and the "gray literature" (project reports available online) between 2000 and 2007 for reports on the proportion of adult patients retained (i.e., remaining in care and on ART) after 6 mo or longer in sub-Saharan African, non-research ART programs, with and without donor support. Estimated retention rates at 6, 12, and 24 mo were calculated and plotted for each program. Retention was also estimated using Kaplan-Meier curves. In sensitivity analyses we considered best-case, worst-case, and midpoint scenarios for retention at 2 y; the best-case scenario assumed no further attrition beyond that reported, while the worst-case scenario assumed that attrition would continue in a linear fashion. We reviewed 32 publications reporting on 33 patient cohorts (74,192 patients, 13 countries). For all studies, the weighted average follow-up period reported was 9.9 mo, after which 77.5% of patients were retained. Loss to follow-up and death accounted for 56% and 40% of attrition, respectively. Weighted mean retention rates as reported were 79.1%, 75.0% and 61.6 % at 6, 12, and 24 mo, respectively. Of those reporting 24 mo of follow-up, the best program retained 85% of patients and the worst retained 46%. Attrition was higher in studies with shorter reporting periods, leading to monthly weighted mean attrition rates of 3.3%/mo, 1.9%/mo, and 1.6%/month for studies reporting to 6, 12, and 24 months, respectively, and suggesting that overall patient retention may be overestimated in the published reports. In sensitivity analyses, estimated retention rates ranged from 24% in the worse case to 77% in the best case at the end of 2 y, with a plausible midpoint scenario of 50%. CONCLUSIONS: Since the inception of large-scale ART access early in this decade, ART programs in Africa have retained about 60% of their patients at the end of 2 y. Loss to follow-up is the major cause of attrition, followed by death. Better patient tracing procedures, better understanding of loss to follow-up, and earlier initiation of ART to reduce mortality are needed if retention is to be improved. Retention varies widely across programs, and programs that have achieved higher retention rates can serve as models for future improvements. [Abstract/Link to Full Text]

Vandenbroucke JP, von Elm E, Altman DG, Gřtzsche PC, Mulrow CD, Pocock SJ, Poole C, Schlesselman JJ, Egger M
Strengthening the Reporting of Observational Studies in Epidemiology (STROBE): explanation and elaboration.
PLoS Med. 2007 Oct 16;4(10):e297.
Much medical research is observational. The reporting of observational studies is often of insufficient quality. Poor reporting hampers the assessment of the strengths and weaknesses of a study and the generalisability of its results. Taking into account empirical evidence and theoretical considerations, a group of methodologists, researchers, and editors developed the Strengthening the Reporting of Observational Studies in Epidemiology (STROBE) recommendations to improve the quality of reporting of observational studies. The STROBE Statement consists of a checklist of 22 items, which relate to the title, abstract, introduction, methods, results and discussion sections of articles. Eighteen items are common to cohort studies, case-control studies and cross-sectional studies and four are specific to each of the three study designs. The STROBE Statement provides guidance to authors about how to improve the reporting of observational studies and facilitates critical appraisal and interpretation of studies by reviewers, journal editors and readers. This explanatory and elaboration document is intended to enhance the use, understanding, and dissemination of the STROBE Statement. The meaning and rationale for each checklist item are presented. For each item, one or several published examples and, where possible, references to relevant empirical studies and methodological literature are provided. Examples of useful flow diagrams are also included. The STROBE Statement, this document, and the associated Web site ( should be helpful resources to improve reporting of observational research. [Abstract/Link to Full Text]

von Elm E, Altman DG, Egger M, Pocock SJ, Gřtzsche PC, Vandenbroucke JP
The Strengthening the Reporting of Observational Studies in Epidemiology (STROBE) statement: guidelines for reporting observational studies.
PLoS Med. 2007 Oct 16;4(10):e296.
Much biomedical research is observational. The reporting of such research is often inadequate, which hampers the assessment of its strengths and weaknesses and of a study's generalisability. The Strengthening the Reporting of Observational Studies in Epidemiology (STROBE) Initiative developed recommendations on what should be included in an accurate and complete report of an observational study. We defined the scope of the recommendations to cover three main study designs: cohort, case-control, and cross-sectional studies. We convened a 2-day workshop in September 2004, with methodologists, researchers, and journal editors to draft a checklist of items. This list was subsequently revised during several meetings of the coordinating group and in e-mail discussions with the larger group of STROBE contributors, taking into account empirical evidence and methodological considerations. The workshop and the subsequent iterative process of consultation and revision resulted in a checklist of 22 items (the STROBE Statement) that relate to the title, abstract, introduction, methods, results, and discussion sections of articles. 18 items are common to all three study designs and four are specific for cohort, case-control, or cross-sectional studies. A detailed Explanation and Elaboration document is published separately and is freely available on the Web sites of PLoS Medicine, Annals of Internal Medicine, and Epidemiology. We hope that the STROBE Statement will contribute to improving the quality of reporting of observational studies. [Abstract/Link to Full Text]

Gong Y, Somwar R, Politi K, Balak M, Chmielecki J, Jiang X, Pao W
Induction of BIM is essential for apoptosis triggered by EGFR kinase inhibitors in mutant EGFR-dependent lung adenocarcinomas.
PLoS Med. 2007 Oct 9;4(10):e294.
BACKGROUND: Mutations in the epidermal growth factor receptor (EGFR) gene are associated with increased sensitivity of lung cancers to kinase inhibitors like erlotinib. Mechanisms of cell death that occur after kinase inhibition in these oncogene-dependent tumors have not been well delineated. We sought to improve understanding of this process in order to provide insight into mechanisms of sensitivity and/or resistance to tyrosine kinase inhibitors and to uncover new targets for therapy. METHODS AND FINDINGS: Using a panel of human lung cancer cell lines that harbor EGFR mutations and a variety of biochemical, molecular, and cellular techniques, we show that EGFR kinase inhibition in drug-sensitive cells provokes apoptosis via the intrinsic pathway of caspase activation. The process requires induction of the proapoptotic BH3-only BCL2 family member BIM (i.e., BCL2-like 11, or BCL2L11); erlotinib dramatically induces BIM levels in sensitive but not in resistant cell lines, and knockdown of BIM expression by RNA interference virtually eliminates drug-induced cell killing in vitro. BIM status is regulated at both transcriptional and posttranscriptional levels and is influenced by the extracellular signal-regulated kinase (ERK) signaling cascade downstream of EGFR. Consistent with these findings, lung tumors and xenografts from mice bearing mutant EGFR-dependent lung adenocarcinomas display increased concentrations of Bim after erlotinib treatment. Moreover, an inhibitor of antiapoptotic proteins, ABT-737, enhances erlotinib-induced cell death in vitro. CONCLUSIONS: In drug-sensitive EGFR mutant lung cancer cells, induction of BIM is essential for apoptosis triggered by EGFR kinase inhibitors. This finding implies that the intrinsic pathway of caspase activation may influence sensitivity and/or resistance of EGFR mutant lung tumor cells to EGFR kinase inhibition. Manipulation of the intrinsic pathway could be a therapeutic strategy to enhance further the clinical outcomes of patients with EGFR mutant lung tumors. [Abstract/Link to Full Text]

Klenerman P, Kim A
HCV-HIV coinfection: simple messages from a complex disease.
PLoS Med. 2007 Oct 9;4(10):e240. [Abstract/Link to Full Text]

Kuipers S, Cannegieter SC, Middeldorp S, Robyn L, Büller HR, Rosendaal FR
The absolute risk of venous thrombosis after air travel: a cohort study of 8,755 employees of international organisations.
PLoS Med. 2007 Sep 25;4(9):e290.
BACKGROUND: The risk of venous thrombosis is approximately 2- to 4-fold increased after air travel, but the absolute risk is unknown. The objective of this study was to assess the absolute risk of venous thrombosis after air travel. METHODS AND FINDINGS: We conducted a cohort study among employees of large international companies and organisations, who were followed between 1 January 2000 and 31 December 2005. The occurrence of symptomatic venous thrombosis was linked to exposure to air travel, as assessed by travel records provided by the companies and organisations. A long-haul flight was defined as a flight of at least 4 h and participants were considered exposed for a postflight period of 8 wk. A total of 8,755 employees were followed during a total follow-up time of 38,910 person-years (PY). The total time employees were exposed to a long-haul flight was 6,872 PY. In the follow-up period, 53 thromboses occurred, 22 of which within 8 wk of a long-haul flight, yielding an incidence rate of 3.2/1,000 PY, as compared to 1.0/1,000 PY in individuals not exposed to air travel (incidence rate ratio 3.2, 95% confidence interval 1.8-5.6). This rate was equivalent to a risk of one event per 4,656 long-haul flights. The risk increased with exposure to more flights within a short time frame and with increasing duration of flights. The incidence was highest in the first 2 wk after travel and gradually decreased to baseline after 8 wk. The risk was particularly high in employees under age 30 y, women who used oral contraceptives, and individuals who were particularly short, tall, or overweight. CONCLUSIONS: The risk of symptomatic venous thrombosis after air travel is moderately increased on average, and rises with increasing exposure and in high-risk groups. [Abstract/Link to Full Text]

Brown M
Defining human differences in biomedicine.
PLoS Med. 2007 Sep 25;4(9):e288. [Abstract/Link to Full Text]

Ellison GT, Smart A, Tutton R, Outram SM, Ashcroft R, Martin P
Racial categories in medicine: a failure of evidence-based practice?
PLoS Med. 2007 Sep 25;4(9):e287. [Abstract/Link to Full Text]

Braun L, Fausto-Sterling A, Fullwiley D, Hammonds EM, Nelson A, Quivers W, Reverby SM, Shields AE
Racial categories in medical practice: how useful are they?
PLoS Med. 2007 Sep 25;4(9):e271. [Abstract/Link to Full Text]

Sismondo S
Ghost management: how much of the medical literature is shaped behind the scenes by the pharmaceutical industry?
PLoS Med. 2007 Sep 25;4(9):e286. [Abstract/Link to Full Text]

Kulkarni GS, Finelli A, Fleshner NE, Jewett MA, Lopushinsky SR, Alibhai SM
Optimal management of high-risk T1G3 bladder cancer: a decision analysis.
PLoS Med. 2007 Sep 25;4(9):e284.
BACKGROUND: Controversy exists about the most appropriate treatment for high-risk superficial (stage T1; grade G3) bladder cancer. Immediate cystectomy offers the best chance for survival but may be associated with an impaired quality of life compared with conservative therapy. We estimated life expectancy (LE) and quality-adjusted life expectancy (QALE) for both of these treatments for men and women of different ages and comorbidity levels. METHODS AND FINDINGS: We evaluated two treatment strategies for high-risk, T1G3 bladder cancer using a decision-analytic Markov model: (1) Immediate cystectomy with neobladder creation versus (2) conservative management with intravesical bacillus Calmette-Guérin (BCG) and delayed cystectomy in individuals with resistant or progressive disease. Probabilities and utilities were derived from published literature where available, and otherwise from expert opinion. Extensive sensitivity analyses were conducted to identify variables most likely to influence the decision. Structural sensitivity analyses modifying the base case definition and the triggers for cystectomy in the conservative therapy arm were also explored. Probabilistic sensitivity analysis was used to assess the joint uncertainty of all variables simultaneously and the uncertainty in the base case results. External validation of model outputs was performed by comparing model-predicted survival rates with independent published literature. The mean LE of a 60-y-old male was 14.3 y for immediate cystectomy and 13.6 y with conservative management. With the addition of utilities, the immediate cystectomy strategy yielded a mean QALE of 12.32 y and remained preferred over conservative therapy by 0.35 y. Worsening patient comorbidity diminished the benefit of early cystectomy but altered the LE-based preferred treatment only for patients over age 70 y and the QALE-based preferred treatment for patients over age 65 y. Sensitivity analyses revealed that patients over the age of 70 y or those strongly averse to loss of sexual function, gastrointestinal dysfunction, or life without a bladder have a higher QALE with conservative therapy. The results of structural or probabilistic sensitivity analyses did not change the preferred treatment option. Model-predicted overall and disease-specific survival rates were similar to those reported in published studies, suggesting external validity. CONCLUSIONS: Our model is, to our knowledge, the first of its kind in bladder cancer, and demonstrated that younger patients with high-risk T1G3 bladder had a higher LE and QALE with immediate cystectomy. The decision to pursue immediate cystectomy versus conservative therapy should be based on discussions that consider patient age, comorbid status, and an individual's preference for particular postcystectomy health states. Patients over the age of 70 y or those who place high value on sexual function, gastrointestinal function, or bladder preservation may benefit from a more conservative initial therapeutic approach. [Abstract/Link to Full Text]

Allan GM, Lexchin J, Wiebe N
Physician awareness of drug cost: a systematic review.
PLoS Med. 2007 Sep 25;4(9):e283.
BACKGROUND: Pharmaceutical costs are the fastest-growing health-care expense in most developed countries. Higher drug costs have been shown to negatively impact patient outcomes. Studies suggest that doctors have a poor understanding of pharmaceutical costs, but the data are variable and there is no consistent pattern in awareness. We designed this systematic review to investigate doctors' knowledge of the relative and absolute costs of medications and to determine the factors that influence awareness. METHODS AND FINDINGS: Our search strategy included The Cochrane Library, EconoLit, EMBASE, and MEDLINE as well as reference lists and contact with authors who had published two or more articles on the topic or who had published within 10 y of the commencement of our review. Studies were included if: either doctors, trainees (interns or residents), or medical students were surveyed; there were more than ten survey respondents; cost of pharmaceuticals was estimated; results were expressed quantitatively; there was a clear description of how authors defined "accurate estimates"; and there was a description of how the true cost was determined. Two authors reviewed each article for eligibility and extracted data independently. Cost accuracy outcomes were summarized, but data were not combined in meta-analysis because of extensive heterogeneity. Qualitative data related to physicians and drug costs were also extracted. The final analysis included 24 articles. Cost accuracy was low; 31% of estimates were within 20% or 25% of the true cost, and fewer than 50% were accurate by any definition of cost accuracy. Methodological weaknesses were common, and studies of low methodological quality showed better cost awareness. The most important factor influencing the pattern and accuracy of estimation was the true cost of therapy. High-cost drugs were estimated more accurately than inexpensive ones (74% versus 31%, Chi-square p < 0.001). Doctors consistently overestimated the cost of inexpensive products and underestimated the cost of expensive ones (binomial test, 89/101, p < 0.001). When asked, doctors indicated that they want cost information and feel it would improve their prescribing but that it is not accessible. CONCLUSIONS: Doctors' ignorance of costs, combined with their tendency to underestimate the price of expensive drugs and overestimate the price of inexpensive ones, demonstrate a lack of appreciation of the large difference in cost between inexpensive and expensive drugs. This discrepancy in turn could have profound implications for overall drug expenditures. Much more focus is required in the education of physicians about costs and the access to cost information. Future research should focus on the accessibility and reliability of medical cost information and whether the provision of this information is used by doctors and makes a difference to physician prescribing. Additionally, future work should strive for higher methodological standards to avoid the biases we found in the current literature, including attention to the method of assessing accuracy that allows larger absolute estimation ranges for expensive drugs. [Abstract/Link to Full Text]

Souza R, Yasuda S, Cristofani S
Treating schizophrenia with DOTS in developing countries: one size does not fit all.
PLoS Med. 2007 Sep 25;4(9):e281; author reply e285. [Abstract/Link to Full Text]

Patel V, Farooq S, Thara R
What is the best approach to treating schizophrenia in developing countries?
PLoS Med. 2007 Jun;4(6):e159.
BACKGROUND TO THE DEBATE: Schizophrenia affects an estimated 25 million people in low- and middle-income countries, with an average lifetime risk of about 1%. The illness is associated with excess mortality from a variety of causes. A 2001 Institute of Medicine report on mental illness in developing countries found that in 1990, over two-thirds of people with schizophrenia in these countries were not receiving any treatment ( The report found no evidence that the proportion of treated people in the developing world had increased since 1990. There is now a debate among mental health professionals in low-income countries over how best to improve patient care. In this article, three psychiatrists give their different viewpoints on the current status of treatment efforts for schizophrenia in the developing world and the measures that can be taken to increase the proportion of patients receiving treatment. [Abstract/Link to Full Text]

Kasi PM, Kassi M, Khawar T
Excessive work hours of physicians in training: maladaptive coping strategies.
PLoS Med. 2007 Sep 25;4(9):e279. [Abstract/Link to Full Text]

Fernández Taylor KR
Excessive work hours of physicians in training in El Salvador: putting patients at risk.
PLoS Med. 2007 Jul;4(7):e205. [Abstract/Link to Full Text]

Recent Articles in BMC Pharmacology

Ranna M, Sinkkonen ST, Möykkynen T, Uusi-Oukari M, Korpi ER
Impact of epsilon and theta subunits on pharmacological properties of alpha3beta1 GABAA receptors expressed in Xenopus oocytes.
BMC Pharmacol. 2006;61.
BACKGROUND: Gamma-aminobutyric acid type A (GABAA) receptors provide the main inhibitory control in the brain. Their heterogeneity may make it possible to precisely target drug effects to selected neuronal populations. In situ hybridization using rat brain sections has revealed a unique expression of GABAA receptor epsilon and theta subunit transcripts in the locus coeruleus, where they are accompanied at least by alpha3, alpha2, beta1 and beta3 subunits. Here, we studied the pharmacology of the human alpha3beta1, alpha3beta1epsilon, alpha3beta1theta and alpha3beta1epsilontheta receptor subtypes expressed in Xenopus oocytes and compared them with the gamma2 subunit-containing receptors. RESULTS: The GABA sensitivites and effects of several positive modulators of GABAA receptors were studied in the absence and the presence of EC25 GABA using the two-electrode voltage-clamp method. We found 100-fold differences in GABA sensitivity between the receptors, alpha3beta1epsilon subtype being the most sensitive and alpha3beta1gamma2 the least sensitive. Also gaboxadol dose-response curves followed the same sensitivity rank order, with EC50 values being 72 and 411 microM for alpha3beta1epsilon and alpha3beta1gamma2 subtypes, respectively. In the presence of EC25 GABA, introduction of the epsilon subunit to the receptor complex resulted in diminished modulatory effects by etomidate, propofol, pregnanolone and flurazepam, but not by pentobarbital. Furthermore, the alpha3beta1epsilon subtype displayed picrotoxin-sensitive spontaneous activity. The theta subunit-containing receptors were efficiently potentiated by the anesthetic etomidate, suggesting that theta subunit could bring the properties of beta2 or beta3 subunits to the receptor complex. CONCLUSION: The epsilon and theta subunits bring additional features to alpha3beta1 GABAA receptors. These receptor subtypes may constitute as novel drug targets in selected brain regions, e.g., in the brainstem locus coeruleus nuclei. [Abstract/Link to Full Text]

Tirkey N, Kaur G, Vij G, Chopra K
Curcumin, a diferuloylmethane, attenuates cyclosporine-induced renal dysfunction and oxidative stress in rat kidneys.
BMC Pharmacol. 2005;515.
BACKGROUND: In India, Curcumin (CMN) is popularly known as "Haldi", and has been well studied due to its economic importance. Traditional Indian medicine claims the use of its powder against biliary disorders, anorexia, coryza, cough, diabetic wounds, hepatic disorder, rheumatism and sinusitis. This study was designed to examine the possible beneficial effect of CMN in preventing the acute renal failure and related oxidative stress caused by chronic administration of cyclosporine (CsA) in rats. CMN was administered concurrently with CsA (20 mg/kg/day s.c) for 21 days. Oxidative stress in kidney tissue homogenates was estimated using thiobarbituric acid reactive substances (TBARS), reduced glutathione (GSH) content, superoxide dismutase (SOD), and Catalase (CAT). Nitrite levels were estimated in serum and tissue homogenates. RESULTS: CsA administration for 21 days produced elevated levels of TBARS and marked depletion of renal endogenous antioxidant enzymes and deteriorated the renal function as assessed by increased serum creatinine, Blood Urea Nitrogen (BUN) and decreased creatinine and urea clearance as compared to vehicle treated rats. CMN markedly reduced elevated levels of TBARS, significantly attenuated renal dysfunction increased the levels of antioxidant enzymes in CsA treated rats and normalized the altered renal morphology. CONCLUSION: In conclusion our study showed that CMN through its antioxidant activity effectively salvaged CsA nephrotoxicity. [Abstract/Link to Full Text]

Angelico P, Velasco C, Guarneri L, Sironi G, Leonardi A, Testa R
Urodynamic effects of oxybutynin and tolterodine in conscious and anesthetized rats under different cystometrographic conditions.
BMC Pharmacol. 2005;514.
BACKGROUND: Antimuscarinic agents are the most popular treatment for overactive bladder and their efficacy in man is well documented, producing decreased urinary frequency and an increase in bladder capacity. During cystometry in rats, however, the main effect reported after acute treatment with antimuscarinics is a decrease in peak micturition pressure together with little or no effect on bladder capacity. In the present experiments we studied the effects, in rats, of the two most widely used antimuscarinic drugs, namely oxybutynin and tolterodine, utilising several different cystometrographic conditions. The aim was to determine the experimental conditions required to reproduce the clinical pharmacological effects of antimuscarinic agents, as seen in humans, in particular their ability to increase bladder capacity. RESULTS: Intravenous or oral administration of tolterodine or oxybutynin in conscious rats utilized 1 day after catheter implantation and with saline infusion at constant rate of 0.1 ml/min, gave a dose-dependent decrease of micturition pressure (MP) with no significant change in bladder volume capacity (BVC). When the saline infusion rate into the bladder was decreased to 0.025 ml/min, the effect of oral oxybutynin was similar to that obtained with the higher infusion rate. Also, experiments were performed in rats in which bladders were infused with suramin (3 and 10 microM) in order to block the non-adrenergic, non-cholinergic component of bladder contraction. Under these conditions, oral administration of oxybutynin significantly reduced MP (as observed previously), but again BVC was not significantly changed. In conscious rats with bladders infused with diluted acetic acid, both tolterodine and oxybutynin administered at the same doses as in animals infused with saline, reduced MP, although the reduction appeared less marked, with no effect on BVC. In conscious rats utilized 5 days after catheter implantation, a situation where inflammation due to surgery is reduced, the effect of tolterodine (i.v.) and oxybutynin (p.o.) on MP was smaller and similar, respectively, to that observed in rats utilized 1 day after catheter implantation, but the increase of BVC was not statistically significant. In anesthetized rats, i.v. administration of oxybutynin again induced a significant decrease in MP, although it was of questionable relevance. Both BVC and threshold pressure were not significantly reduced. The number and amplitude of high frequency oscillations in MP were unmodified by treatment. Finally, in conscious obstructed rats, intravenous oxybutynin did not modify frequency and amplitude of non-voiding contractions or bladder capacity and micturition volume. CONCLUSION: Despite the different experimental conditions used, the only effect on cystometrographic parameters of oxybutynin and tolterodine in anesthetized and conscious rats was a decrease in MP, whereas BVC was hardly and non-significantly affected. Therefore, it is difficult to reproduce in rats the cystometrographic increase in BVC as observed in humans after chronic administration of antimuscarinic agents, whereas the acute effects seem more similar. [Abstract/Link to Full Text]

Howell G, West L, Jenkins C, Lineberry B, Yokum D, Rockhold R
In vivo antimuscarinic actions of the third generation antihistaminergic agent, desloratadine.
BMC Pharmacol. 2005;513.
BACKGROUND: Muscarinic receptor mediated adverse effects, such as sedation and xerostomia, significantly hinder the therapeutic usefulness of first generation antihistamines. Therefore, second and third generation antihistamines which effectively antagonize the H1 receptor without significant affinity for muscarinic receptors have been developed. However, both in vitro and in vivo experimentation indicates that the third generation antihistamine, desloratadine, antagonizes muscarinic receptors. To fully examine the in vivo antimuscarinic efficacy of desloratadine, two murine and two rat models were utilized. The murine models sought to determine the efficacy of desloratadine to antagonize muscarinic agonist induced salivation, lacrimation, and tremor. Desloratadine's effect on the cardiovascular system was explored in both rodent models. RESULTS: In the pithed rat, both desloratadine (1.0 mg/kg, i.v.) and the muscarinic M2 selective antagonist, methoctramine (0.5 mg/kg, i.v.), inhibited negative inotropic (left ventricular dP/dt) effects caused by oxotremorine, a nonselective muscarinic agonist (p < 0.05). Negative chronotropic effects caused by oxotremorine were inhibited by desloratadine, methoctramine, and the muscarinic M3 selective antagonist, 4-DAMP (1.0 mg/kg, i.v.). A late positive inotropic event observed after the initial decrease was inhibited by all three test compounds with desloratadine and 4-DAMP being the most efficacious. In the conscious animal, inhibition of baroreflex-mediated bradycardia was evaluated. Unlike atropine (0.5 mg/kg, i.v.), desloratadine did not alter this bradycardia. The antimuscarinic action of desloratadine on salivation, lacrimation, and tremor was also explored. In urethane-anesthetized (1.5 g/kg, i.p.) male ICR mice (25-35 g) desloratadine (1.0, 5.0 mg/kg) did not inhibit oxotremorine-induced (0.5 mg/kg, s.c.) salivation, unlike atropine (0.5 mg/kg) and 4-DAMP (1.0 mg/kg). In conscious mice, desloratadine failed to inhibit oxotremorine-induced (0.5 mg/kg, s.c.) salivation, lacrimation, and tremor. However, desloratadine did inhibit oxotremorine-induced tremor in phenylephrine pretreated animals. CONCLUSION: The presented data demonstrate that the third generation antihistamine, desloratadine, does not significantly antagonize peripheral muscarinic receptors mediating salivation and lacrimation, therefore, xerostomia and dry eyes should not be observed with therapeutic use of desloratadine. Our data also indicate when administered to a patient with a compromised blood-brain barrier, desloratadine may cause sedation. Patients with compromised cardiovascular systems should be closely monitored when administered desloratadine based on our results that desloratadine has the ability to interfere with normal cardiovascular function mediated by muscarinic receptors. [Abstract/Link to Full Text]

Levine L
Tetrandrine and thapsigargin release arachidonic acid from cells in culture and stimulate prostacyclin production in rat liver cells, but may do so by different pathways.
BMC Pharmacol. 2005;5(1):12.
BACKGROUND: Tetrandrine inhibits tumor cell proliferation and demonstrates chemoprevention in cancer models. Speculation on the association between its effects on K+ and Ca2+ channels and cancer chemoprevention has been made. Thapsigargin also affects K+ and Ca2+ conductance. Thapsigargin, however, is a weak tumor promoter in the two-stage model of mouse skin carcinogenesis, yet it can induce apoptosis in androgen-independent prostatic cancer cells. I have postulated that arachidonic acid release from cells in culture is associated with cancer chemoprevention. The effects of tetrandrine and thapsigargin on arachidonic acid release from human colon carcinoma and rat liver cells and prostacyclin production by rat liver cells are compared in the current studies. RESULTS: Tetrandrine and thapsigargin stimulate arachidonic acid release from human colon carcinoma and rat liver cells and prostacyclin production in rat liver cells. The stimulation by tetrandrine is not affected by incubation with actinomycin D, 100 mM KCl, the [Ca2+]i chelator, 1,2-bis (o-amino-5-fluorophenoxy) ethane-N,N,N',N',-tetraacetic acid tetraacetoxymethylester (BAPTA/AM) or in the absence of extracellular Ca2+. In contrast, stimulation by thapsigargin is inhibited by incubation with actinomycin D, 100 mM KCl, BAPTA/AM or in the absence of extracellular Ca2+. CONCLUSION: Both tetrandrine and thapsigargin stimulate arachidonic acid release, but based on the different results obtained in the presence of actinomycin D, the [Ca2+]i chelator, 100 mM KCl and in the absence of extracellular Ca2+, the mechanisms leading to this release and pathways leading to apoptosis and/or cancer chemoprevention may be different. Stimulations by tetrandrine may be mediated by activation of a secretory phospholipase A2, whereas thapsigargin's stimulations may be mediated by the cytoplasmic Ca2+-dependent phospholipase A2. [Abstract/Link to Full Text]

Monfort P, Felipo V
Long-term potentiation in hippocampus involves sequential activation of soluble guanylate cyclase, cGMP-dependent protein kinase and cGMP-degrading phosphodiesterase, alterations in hyperammonemia.
BMC Pharmacol. 2005;5 Suppl 1P66. [Abstract/Link to Full Text]

Lu DY, Huang M, Xu CH, Yang WY, Hu CX, Lin LP, Tong LJ, Li MH, Lu W, Zhang XW, Ding J
Anti-proliferative effects, cell cycle G2/M phase arrest and blocking of chromosome segregation by probimane and MST-16 in human tumor cell lines.
BMC Pharmacol. 2005;5(1):11.
BACKGROUND: Anticancer bisdioxopiperazines, including ICRF-154, razoxane (Raz, ICRF-159) and ICRF-193, are a family of anticancer agents developed in the UK, especially targeting metastases of neoplasms. Two other bisdioxopiperazine derivatives, probimane (Pro) and MST-16, were synthesized at the Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, China. Cytotoxic activities and mechanisms of Raz (+)-steroisomer (ICRF-187, dexrazoxane), Pro and MST-16 against tumor cells were evaluated by MTT colorimetry, flow cytometry and karyotyping. RESULTS: Pro was cytotoxic to human tumor cell lines in vitro (IC50<50 microM for 48 h). Four human tumor cell lines (SCG-7901, K562, A549 and HL60) were susceptible to Pro at low inhibitory concentrations (IC50 values < 10 microM for 48 h). Although the IC50 against HeLa cell line of vincristine (VCR, 4.56 microM), doxorubicin (Dox, 1.12 microM) and 5-fluoruouracil (5-Fu, 0.232 microM) are lower than Pro (5.12 microM), ICRF-187 (129 microM) and MST-16 (26.4 microM), VCR, Dox and 5-Fu shows a low dose-related - high cytotoxic activity. Time-response studies showed that the cytotoxic effects of Pro are increased for 3 days in human tumor cells, whereas VCR, Dox and 5-Fu showed decreased cytotoxic action after 24 h. Cell cycle G2/M phase arrest and chromosome segregation blocking by Pro and MST-16 were noted. Although there was similar effects of Pro and MST-16 on chromosome segregation blocking action and cell cycle G2/M phase arrest at 1- 4 microM, cytotoxicity of Pro against tumor cells was higher than that of MST-16 in vitro by a factor of 3- 10 folds. Our data show that Pro may be more effective against lung cancer and leukemia while ICRF-187 and MST-16 shows similar IC50 values only against leukemia. CONCLUSION: It suggests that Pro has a wider spectrum of cytotoxic effects against human tumor cells than other bisdioxopiperazines, especially against solid tumors, and with a single cytotoxic pathway of Pro and MST-16 affecting chromosome segregation and leading also to cell G2/ M phase arrests, which finally reduces cell division rates. Pro may be more potent than MST-16 in cytotoxicity. High dose- and time- responses of Pro, when compared with VCR, 5-Fu and Dox, were seen that suggest a selectivity of Pro against tumor growth. Compounds of bisdioxopiperazines family may keep up their cytotoxic effects longer than many other anticancer drugs. [Abstract/Link to Full Text]

Hassan MA, Ketat AF
Sildenafil citrate increases myocardial cGMP content in rat heart, decreases its hypertrophic response to isoproterenol and decreases myocardial leak of creatine kinase and troponin T.
BMC Pharmacol. 2005;5(1):10.
BACKGROUND: Cardiac hypertrophy is a major risk factor for morbidity and mortality in a number of cardiovascular diseases. Consequently, the signaling pathways that inhibit cardiac hypertrophy are currently receiving much interest. Among them, nitric oxide (NO), signaling via cGMP and cGMP-dependent protein kinase I, has been recognized as a negative regulator of cardiac hypertrophy. The present study investigated the in-vivo effect of sildenafil as a phosphodiestrase-5A (PDE-5A) inhibitor on the hypertrophic response of rat heart to isoproterenol and the relation of this effect to the level of myocardial cGMP and integrity of the constitutive nitric oxide synthase (cNOS) activity. RESULTS: The results showed that daily intraperitoneal administration of sildenafil per se for 10 days was without noticeable adverse effects on survival or myocardium. Conversely, daily subcutaneous administration of isoproterenol for 10 days caused significant myocardial hypertrophy, cell injury and decline in survival. When sildenafil was injected daily, one hour before isoproterenol, survival was significantly improved and the myocardium didn't show significant hypertrophy or cell injury. Interestingly, sildenafil was accompanied by significant rise in myocardial cGMP level, a parameter which was found in the present study to possess a significant negative correlation with cardiac hypertrophy and leak of cardiac troponin T into serum. At the same time, cGMP was found to possess a positive correlation with myocardial creatine kinase activity that reflects the efficiency of the energy utilization processes in the myocardium. However, in rats given Nomega-nitro-L-arginine (L-NNA) as a competitive inhibitor of cNOS, sildenafil failed to show any favorable effect on survival or the myocardial injury parameters used to assess isoproterenol-induced injury. CONCLUSION: The present study suggests that increased cardiac cGMP level by sildenafil have a cardioprotective effect probably through acting as a post-receptor negative regulator of cardiac sympathetic responsiveness. Integrity of NOS function was an essential prerequisite for sildenafil's mediated cardioprotection encountered in the present study. [Abstract/Link to Full Text]

Ward GR, Abdel-Rahman AA
Effect of testosterone replacement or duration of castration on baroreflex bradycardia in conscious rats.
BMC Pharmacol. 2005;5(1):9.
BACKGROUND: In this study, we tested the hypothesis that 17beta-estradiol contributes to testosterone-mediated restoration of baroreflex-mediated bradycardia in short-term (3 weeks) castrated rats. Further, a reported increase in serum testosterone after long-term (6 weeks) castration constituted a basis for testing the hypothesis that a spontaneous increase in serum testosterone or androstenedione in this model causes a commensurate increase in baroreflex-mediated bradycardia. RESULTS: Testosterone (1 week) replacement enhanced baroreflex-mediated bradycardia in short-term castrated rats without changing 17beta-estradiol level. A spontaneous recovery of baroreflex-mediated bradycardia occurred following long-term castration, although circulating testosterone and androstenedione remained suppressed. CONCLUSION: The data suggest: 1) 17beta-Estradiol does not contribute to testosterone restoration of the baroreflex-mediated bradycardia in short-term castrated rats. 2) The long-term modulation of baroreflex-mediated bradycardia occurs independent of androgens, or the baroreflex mechanism may become adapted to low levels of circulating androgens. [Abstract/Link to Full Text]

Liu H, Farley JM
Effects of first and second generation antihistamines on muscarinic induced mucus gland cell ion transport.
BMC Pharmacol. 2005;5(1):8.
BACKGROUND: The first generation antihistamines, such as diphenhydramine, are fairly potent muscarinic antagonists in addition to being H1 selective antihistamines. The antimuscarinic action is often not desirable since it is in part responsible for the drying of secretions in the airways and the sedative effect. We therefore examined a number of antihistamines for antimuscarinic effects on ion transport by mucus gland cells isolated from the airways of swine. Enzymatically isolated airway mucus gland cells were purified utilizing density gradients and grown in culture on porous inserts (Millicell HA) at an air interface. Cells grown in this manner maintain phenotype and polarity. Transport of ions, as short-circuit current measured under voltage-clamp, was measured in response to acetylcholine (ACh) or histamine applied to the serosal side of the gland cell layers. Concentration-response relationships for ACh or histamine were generated in the presence and absence of various drugs. The potencies against muscarinic receptor activation were estimated using the dose-ratio method of Schild. RESULTS: Three known muscarinic antagonists were used to validate the system. Atropine had a pA2 of 9.4 +/- 0.1 (n = 9). 4-DAMP and methoctramine had pA2 values of 8.6 +/- 0.1 and 5.6 +/- 0.1, respectively (n = 12, 11) all consistent with inhibition of an M3 subtype muscarinic receptor. The rank order of potency of the antihistamines against the inhibition of M3 receptors was desloratadine = diphenhydramine > hydroxyzine (pA2; 6.4, 6.2, 4.8, respectively). pA2 values for fexofenadine, loratadine and cetirizine were not determined since they had no effect on the cholinergic response at the highest drug concentrations tested (10, 10 and 100 microM, respectively). The pA2 values for the antihistamines against the histamine response could not be calculated, but the estimates of the rank order of potency were estimated to be desloratadine > cetirizine approximate to hydroxyzine > fexofenadine > loratadine > diphenhydramine. CONCLUSION: The rank order of selectivity for histamine receptors over muscarinic receptors was estimated to be cetirizine approximate to fexofenadine > loratadine > desloratadine > or = hydroxyzine > or = diphenhydramine. [Abstract/Link to Full Text]

Cha MC, Lin A, Meckling KA
Low dose docosahexaenoic acid protects normal colonic epithelial cells from araC toxicity.
BMC Pharmacol. 2005;5(1):7.
BACKGROUND: The nucleoside analogue arabinosylcytosine (araC) has been used for many years in the treatment of acute leukemia. Evidence in the literature suggests that araC may inhibit the growth of human colon carcinoma cell lines as well. Because araC action interferes with normal nucleoside metabolism, it is highly toxic to a number of normal cell types including bone marrow and intestinal mucosa cells. Here we investigate whether the omega-3 fatty acid docosahexaenoic acid (DHA) could selectively target araC toxicity toward colonic tumor cells while protecting the normal cells in vitro. RESULTS: Cultures of normal rat colonic epithelial cells (4D/WT) and those transformed by v-src (D/v-src) were supplemented with graded concentrations of DHA or arachidonic acid (AA) alone or in combination with araC. AraC was only 1.6 fold more toxic to D/v-src than 4D/WT in cultures without added fatty acids. Supplementing with as little as 3 muM of either AA or DHA increased araC toxicity by more than 30-fold in the tumorigenic cells. The toxic effect of araC on the normal cells was also increased by the fatty acid supplementation. IC50 values were decreased 1.7 fold by DHA in the 4D/WT cells but a more than 7-fold decrease was observed during AA supplementation. As a result, the therapeutic index of araC (IC50 normal/IC50 tumor) was more than 3-fold higher in the DHA than the AA supplemented cells. The expression of protein kinase C isoform epsilon was decreased in AA alone supplemented D/v-src cultures but in combination with araC decreased only in DHA supplemented 4D/WT cells. CONCLUSION: Low dose DHA supplementation may enhance araC chemotherapy in colon cancer while protecting normal tissues, possibly through control of PKC signalling pathways. [Abstract/Link to Full Text]

Mayer AM, Hall ML, Lynch SM, Gunasekera SP, Sennett SH, Pomponi SA
Differential modulation of microglia superoxide anion and thromboxane B2 generation by the marine manzamines.
BMC Pharmacol. 2005;5(1):6.
BACKGROUND: Thromboxane B2 (TXB2) and superoxide anion (O2-) are neuroinflammatory mediators that appear to be involved in the pathogenesis of several neurodegenerative diseases. Because activated-microglia are the main source of TXB2 and O2- in these disorders, modulation of their synthesis has been hypothesized as a potential therapeutic approach for neuroinflammatory disorders. Marine natural products have become a source of novel agents that modulate eicosanoids and O2- generation from activated murine and human leukocytes. With the exception of manzamine C, all other manzamines tested are characterized by a complex pentacyclic diamine linked to C-1 of the beta-carboline moiety. These marine-derived alkaloids have been reported to possess a diverse range of bioactivities including anticancer, immunostimulatory, insecticidal, antibacterial, antimalarial and antituberculosis activities. The purpose of this investigation was to conduct a structure-activity relationship study with manzamines (MZ) A, B, C, D, E and F on agonist-stimulated release of TXB2 and O2- from E. coli LPS-activated rat neonatal microglia in vitro. RESULTS: The manzamines differentially attenuated PMA (phorbol 12-myristate 13-acetate)-stimulated TXB2 generation in the following order of decreasing potency: MZA (IC50 < 0.016 microM) > MZD (IC50 = 0.23 microM) > MZB (IC50 = 1.6 microM) > MZC (IC50 = 2.98 microM) > MZE and F (IC50 > 10 microM). In contrast, there was less effect on OPZ (opsonized zymosan)-stimulated TXB2 generation: MZB (IC50 = 1.44 microM) > MZA (IC50 = 3.16 microM) > MZC (IC50 = 3.34 microM) > MZD, MZE and MZF (IC50 > 10 microM). Similarly, PMA-stimulated O2- generation was affected differentially as follows: MZD (apparent IC50 < 0.1 microM) > MZA (IC50 = 0.1 microM) > MZB (IC50 = 3.16 microM) > MZC (IC50 = 3.43 microM) > MZE and MZF (IC50 > 10 microM). In contrast, OPZ-stimulated O2- generation was minimally affected: MZB (IC50 = 4.17 microM) > MZC (IC50 = 9.3 microM) > MZA, MZD, MZE and MZF (IC50 > 10 microM). From the structure-activity relationship perspective, contributing factors to the observed differential bioactivity on TXB2 and O2- generation are the solubility or ionic forms of MZA and D as well as changes such as saturation or oxidation of the beta carboline or 8-membered amine ring. In contrast, the fused 13-membered macrocyclic and isoquinoline ring system, and any substitutions in these rings would not appear to be factors contributing to bioactivity. CONCLUSION: To our knowledge, this is the first experimental study that demonstrates that MZA, at in vitro concentrations that are non toxic to E. coli LPS-activated rat neonatal microglia, potently modulates PMA-stimulated TXB2 and O2- generation. MZA may thus be a lead candidate for the development of novel therapeutic agents for the modulation of TXB2 and O2- release in neuroinflammatory diseases. Marine natural products provide a novel and rich source of chemical diversity that can contribute to the design and development of new and potentially useful anti-inflammatory agents to treat neurodegenerative diseases. [Abstract/Link to Full Text]

Upton RN, Ludbrook GL
Pharmacokinetic-pharmacodynamic modelling of the cardiovascular effects of drugs - method development and application to magnesium in sheep.
BMC Pharmacol. 2005;5(1):5.
BACKGROUND: There have been few reports of pharmacokinetic models that have been linked to models of the cardiovascular system. Such models could predict the cardiovascular effects of a drug under a variety of circumstances. Limiting factors may be the lack of a suitably simple cardiovascular model, the difficulty in managing extensive cardiovascular data sets, and the lack of physiologically based pharmacokinetic models that can account for blood flow changes that may be caused by a drug. An approach for addressing these limitations is proposed, and illustrated using data on the cardiovascular effects of magnesium given intravenously to sheep.The cardiovascular model was based on compartments for venous and arterial blood. Blood flowed from arterial to venous compartments via a passive flow through a systemic vascular resistance. Blood flowed from venous to arterial via a pump (the heart-lung system), the pumping rate was governed by the venous pressure (Frank-Starling mechanism). Heart rate was controlled via the difference between arterial blood pressure and a set point (Baroreceptor control). Constraints were made to pressure-volume relationships, pressure-stroke volume relationships, and physical limits were imposed to produce plausible cardiac function curves and baseline cardiovascular variables. "Cardiovascular radar plots" were developed for concisely displaying the cardiovascular status. A recirculatory kinetic model of magnesium was developed that could account for the large changes in cardiac output caused by this drug. Arterial concentrations predicted by the kinetic model were linked to the systemic vascular resistance and venous compliance terms of the cardiovascular model. The kinetic-dynamic model based on a training data set (30 mmol over 2 min) was used to predict the results for a separate validation data set (30 mmol over 5 min). RESULTS: The kinetic-dynamic model was able to describe the training data set. A recirculatory kinetic model was a good description of the acute kinetics of magnesium in sheep. The volume of distribution of magnesium in the lungs was 0.89 L, and in the body was 4.02 L. A permeability term (0.59 L min-1) described the distribution of magnesium into a deeper (probably intracellular) compartment. The final kinetic-dynamic model was able to predict the validation data set. The mean prediction error for the arterial magnesium concentrations, cardiac output and mean arterial blood pressure for the validation data set were 0.02, 3.0 and 6.1%, respectively. CONCLUSION: The combination of a recirculatory model and a simple two-compartment cardiovascular model was able to describe and predict the kinetics and cardiovascular effects of magnesium in sheep. [Abstract/Link to Full Text]

Lindemalm S, Savic RM, Karlsson MO, Juliusson G, Liliemark J, Albertioni F
Application of population pharmacokinetics to cladribine.
BMC Pharmacol. 2005;5(1):4.
BACKGROUND: The nucleoside analog cladribine is used for the treatment of a variety of indolent B- and T-cell lymphoid malignancies. The primary aim of the study was to evaluate the population distribution of pharmacokinetic parameters in patients undergoing treatment with cladribine and to detect the influence of different covariates on the pharmacokinetic parameters. METHODS: This pharmacokinetic study presents the results of a retrospective population pharmacokinetic analysis based on pooled data from 161 patients, who were given cladribine in different administration routes in various dosing regimens. The plasma concentrations of cladribine were determined by reversed-phase high-performance liquid chromatography using a solid phase extraction with a limit of quantitation of 1 nM using 1 mL of plasma. RESULTS: A three compartment structural model best described the disposition of cladribine. Clearance was found to be 39.3 L/hour, with a large interindividual variability. The half-life for the terminal phase was 16 hours. Bioavailability was 100% and 35% for subcutaneous and oral administration, respectively, with low interindividual variability. None of the investigated covariates were found to be correlated with the pharmacokinetic parameters. CONCLUSION: As interindividual variability in apparent clearance after oral administration was not significantly higher compared to that following infusion, cladribine could be administered orally instead of intravenously if compensated for its lower bioavailability. Individualized dosing on basis of body surface area or weight does not represent an improvement in this study as compared to administering a fixed dose to all patients. [Abstract/Link to Full Text]

Schatter B, Jin S, Löffelholz K, Klein J
Cross-talk between phosphatidic acid and ceramide during ethanol-induced apoptosis in astrocytes.
BMC Pharmacol. 2005 Feb 4;5(1):3.
BACKGROUND: Ethanol inhibits proliferation in astrocytes, an effect that was recently linked to the suppression of phosphatidic acid (PA) formation by phospholipase D (PLD). The present study investigates ethanol's effect on the induction of apoptosis in astrocytes and the formation of ceramide, an apoptotic signal. Evidence is presented that the formation of PA and ceramide may be reciprocally linked during ethanol exposure. RESULTS: In cultured rat cortical astrocytes, ethanol (0.3-1 %, v/v) induced nuclear fragmentation and DNA laddering indicative of apoptosis. Concomitantly, in cells prelabeled with [3H]-serine, ethanol caused a dose-dependent, biphasic increase of the [3H]-ceramide/[3H]-sphingomyelin ratio after 1 and 18 hours of incubation. As primary alcohols such as ethanol and 1-butanol were shown to inhibit the phospholipase D (PLD)-mediated formation of PA, a mitogenic lipid messenger, we tested their effects on ceramide formation. In astrocytes prelabeled with [3H]-serine, ethanol and 1-butanol, in contrast to t-butanol, significantly increased the formation of [3H]-ceramide. Moreover, exogenous PA, added to transiently permeabilized astrocytes, suppressed ethanol-induced [3H]-ceramide formation. Vice versa, addition of C2-ceramide to astrocytes inhibited PLD activity induced by serum or phorbol ester. CONCLUSION: We propose that the formation of ceramide in ethanol-exposed astrocytes is secondary to the disruption of phospholipase D signaling. Ethanol reduces the PA:ceramide ratio in fetal astrocytes, a mechanism which likely participates in ethanol-induced glial apoptosis during brain development. [Abstract/Link to Full Text]

Tirkey N, Pilkhwal S, Kuhad A, Chopra K
Hesperidin, a citrus bioflavonoid, decreases the oxidative stress produced by carbon tetrachloride in rat liver and kidney.
BMC Pharmacol. 2005 Jan 31;5(1):2.
BACKGROUND: CCl4 is a well-established hepatotoxin inducing liver injury by producing free radicals. Exposure to CCl4 also induces acute and chronic renal injuries. The present study was designed to establish the protective effect of hesperidin (HDN), a citrus bioflavonoid, on CCl4-induced oxidative stress and resultant dysfunction of rat liver and kidney. METHODS: Animals were pretreated with HDN (100 and 200 mg/kg orally) for one week and then challenged with CCl4 (2 ml/kg/s.c.) in olive oil. Rats were sacrificed by carotid bleeding under ether anesthesia. Liver enzymes, urea and creatinine were estimated in serum. Oxidative stress in liver and kidney tissue was estimated using Thiobarbituric acid reactive substances (TBARS), glutathione (GSH) content, superoxide dismutase(SOD), and Catalase (CAT) RESULTS: CCl4 caused a marked rise in serum levels of ALT and AST (P < 0.05). TBARS levels were significantly increased whereas GSH, SOD and CAT levels decreased in the liver and kidney homogenates of CCl4 treated rats. HDN (200 mg/kg) successfully attenuated these effects of CCl4 CONCLUSION: In conclusion, our study demonstrated a protective effect of HDN in CCl4 induced oxidative stress in rat liver and kidney. This protective effect of HDN can be correlated to its direct antioxidant effect. [Abstract/Link to Full Text]

Berggard C, Damberg M, Oreland L
Brainstem levels of transcription factor AP-2 in rat are changed after treatment with phenelzine, but not with citalopram.
BMC Pharmacol. 2005 Jan 21;5(1):1.
BACKGROUND: Before therapeutic effect is obtained after treatment with antidepressant drugs, like serotonin selective reuptake inhibitors (SSRIs), tricyclic antidepressants (TCAs) and monoamine oxidase inhibitors (MAO-Is) there is an initial lag-period of a few weeks. Neuronal adaptations on a molecular level are supposed to be involved in the initiation of the antidepressant effect. Transcription factor AP-2 is essential for neuronal development and many genes involved in the brainstem monoaminergic systems have binding sites for AP-2 in their regulatory regions. The genotype of the AP-2beta isoform has been associated with e.g. anxiety-related personality traits and with platelet MAO activity. In addition, previous studies have shown that the levels of AP-2alpha and AP-2beta in rat whole brain were decreased after 10 days of treatment with citalopram (SSRI) and imipramine (TCA), and were increased with phenelzine (MAO-I). RESULTS: In the present study, we report that treatment with citalopram for 1, 7 or 21 days did not have effect on the AP-2 levels in rat brainstem. However, after treatment with phenelzine for 1, 7 or 21 days the levels of AP-2alpha and AP-2beta had increased after 7 days, but had returned to control levels at day 21. CONCLUSION: The decrease in AP-2 levels in rat whole brain previously seen after treatment with citalopram does not seem to be localised to the brainstem, it may rather occur in the monoaminergic terminal projection areas. The present data suggest that the increase in AP-2 levels previously seen in rat whole brain after subchronic treatment with phenelzine is located in the brainstem. It cannot, however, be excluded that other brain regions are involved. [Abstract/Link to Full Text]

Lu DY, Xu B, Ding J
Antitumor effects of two bisdioxopiperazines against two experimental lung cancer models in vivo.
BMC Pharmacol. 2004 Dec 24;4(1):32.
BACKGROUND: Probimane (Pro), an anti-cancer agent originating in China, was derived from razoxane (ICRF-159, Raz), a drug created in Britain, specifically targeting at cancer metastasis and as a cardioprotectant of anthrocyclines. Pro and Raz are bisdioxopiperazine compounds. In this work, we evaluated the anti-tumor and anti-metastatic effects of Pro and Raz in vivo against two lung tumor models, one of murine origin (Lewis lung carcinoma, LLC) and one of human origin (LAX-83). RESULTS: After determining the lethal dosage of Pro and Raz, we assessed and compared the inhibitory effects of Pro and Raz against primary tumor growth and metastatic occurrences of LLC at the dosage of LD5. Pro and Raz were active against primary tumor growth and significantly inhibited pulmonary metastasis of LLC at same dose-ranges (inhibitory rates > 90 %). Both Raz and Pro were effective in 1, 5, and 9 day administration schedules. Three different schedules of Raz and Pro were effective against the primary tumor growth of LLC (35-50 %). The synergistic anticancer effect of Raz with bleomycin (Ble) (from 41.3 % to 73.3 %) was more obvious than those with daunorubicin (Dau) (from 33.1 % to 56.3 %) in the LLC tumor model. Pro was also seen to have synergistic anti-cancer effects with Ble in the LLC model. Both Raz and Pro inhibited the growth of LAX 83 in a statistically significant manner. CONCLUSIONS: These data suggest that both Raz and Pro may have anti-tumor potentiality and Raz and Pro have combinative effects with Ble or Dau. The potential targets of bisdioxopiperazines may include lung cancers, especially on tumor metastasis. The anti-cancer effects of Raz and Pro can be increased with the help of other anticancer drugs. [Abstract/Link to Full Text]

Jensen LH, Dejligbjerg M, Hansen LT, Grauslund M, Jensen PB, Sehested M
Characterisation of cytotoxicity and DNA damage induced by the topoisomerase II-directed bisdioxopiperazine anti-cancer agent ICRF-187 (dexrazoxane) in yeast and mammalian cells.
BMC Pharmacol. 2004 Dec 2;4(1):31.
BACKGROUND: Bisdioxopiperazine anti-cancer agents are inhibitors of eukaryotic DNA topoisomerase II, sequestering this protein as a non-covalent protein clamp on DNA. It has been suggested that such complexes on DNA represents a novel form of DNA damage to cells. In this report, we characterise the cytotoxicity and DNA damage induced by the bisdioxopiperazine ICRF-187 by a combination of genetic and molecular approaches. In addition, the well-established topoisomerase II poison m-AMSA is used for comparison. RESULTS: By utilizing a panel of Saccharomyces cerevisiae single-gene deletion strains, homologous recombination was identified as the most important DNA repair pathway determining the sensitivity towards ICRF-187. However, sensitivity towards m-AMSA depended much more on this pathway. In contrast, disrupting the post replication repair pathway only affected sensitivity towards m-AMSA. Homologous recombination (HR) defective irs1SF chinese hamster ovary (CHO) cells showed increased sensitivity towards ICRF-187, while their sensitivity towards m-AMSA was increased even more. Furthermore, complementation of the XRCC3 deficiency in irs1SF cells fully abrogated hypersensitivity towards both drugs. DNA-PKcs deficient V3-3 CHO cells having reduced levels of non-homologous end joining (NHEJ) showed slightly increased sensitivity to both drugs. While exposure of human small cell lung cancer (SCLC) OC-NYH cells to m-AMSA strongly induced gammaH2AX, exposure to ICRF-187 resulted in much less induction, showing that ICRF-187 generates fewer DNA double strand breaks than m-AMSA. Accordingly, when yeast cells were exposed to equitoxic concentrations of ICRF-187 and m-AMSA, the expression of DNA damage-inducible genes showed higher levels of induction after exposure to m-AMSA as compared to ICRF-187. Most importantly, ICRF-187 stimulated homologous recombination in SPD8 hamster lung fibroblast cells to lower levels than m-AMSA at all cytotoxicity levels tested, showing that the mechanism of action of bisdioxopiperazines differs from that of classical topoisomerase II poisons in mammalian cells. CONCLUSION: Our results point to important differences in the mechanism of cytotoxicity induced by bisdioxopiperazines and topoisomerase II poisons, and suggest that bisdioxopiperazines kill cells by a combination of DNA break-related and DNA break-unrelated mechanisms. [Abstract/Link to Full Text]

Sreemantula S, Boini KM, Nammi S
Reserpine methonitrate, a novel quaternary analogue of reserpine augments urinary excretion of VMA and 5-HIAA without affecting HVA in rats.
BMC Pharmacol. 2004 Nov 16;4(1):30.
BACKGROUND: Reserpine, an alkaloid from Rauwolfia serpentina was widely used for its antihypertensive action in the past. In later years, its use has been reduced because of precipitation of depression and extra pyramidal symptoms due to its central action. In the present investigation, reserpine methonitrate (RMN), a novel quaternary analogue of reserpine was synthesised and evaluated biochemically for its central and peripheral amine depleting actions in rats while its influence on the blood pressure was measured in anaesthetized rats in comparison with reserpine RESULTS: Reserpine treatment (5 mg/kg) produced a significant increase in the urinary excretion of VMA, 5-HIAA and HVA while RMN at doses of equal to and double the equimolar doses of reserpine (5 and 10 mg/kg) produced significant increase in VMA and 5-HIAA excretion without producing any effect on HVA excretion compared to control animals. Reserpine in the dose range of 0.5 to 15 microg/kg produced significant reduction in blood pressure compared to control. RMN was also found to produce significant decrease in blood pressure at doses of 10, 25 and 50 microg/kg body weight in comparison to control. The results indicated peripheral depletion of biogenic amines by RMN without affecting the central stores of the amines. CONCLUSIONS: The present study clearly indicated that the quaternization of reserpine restricts its transfer across the blood-brain barrier and could be the reason for its selective peripheral action. It is also clear that the hypotensive actions of RMN could be due to peripheral depletion of catecholamines. [Abstract/Link to Full Text]

Narang D, Sood S, Thomas MK, Dinda AK, Maulik SK
Effect of dietary palm olein oil on oxidative stress associated with ischemic-reperfusion injury in isolated rat heart.
BMC Pharmacol. 2004 Nov 9;4(1):29.
BACKGROUND: Palm olein oil (PO), obtained from refining of palm oil is rich in monounsaturated fatty acid and antioxidant vitamins and is widely used as oil in diet in many parts of the world including India. Palm oil has been reported to have beneficial effects in oxidative stress associated with hypertension and arterial thrombosis. Oxidative stress plays a major role in the etiopathology of myocardial ischemic-reperfusion injury (IRI) which is a common sequel of ischemic heart disease. Antioxidants have potent therapeutic effects on both ischemic heart disease and ischemic-reperfusion injury. Information on the effect of PO on ischemic-reperfusion injury is, however, lacking. In the present study, the effect of dietary palm olein oil on oxidative stress associated with IRI was investigated in an isolated rat heart model. Wistar rats (150-200 gm) of either sex were divided into three different groups (n = 16). Rats were fed with palm olein oil supplemented commercial rat diet, in two different doses [5% v / w (PO 5) and 10% v / w (PO 10) of diet] for 30 days. Control rats (C) were fed with normal diet. After 30 days, half the rats from each group were subjected to in vitro myocardial IRI (20 min of global ischemia, followed by 40 min of reperfusion). Hearts from all the groups were then processed for biochemical and histopathological studies. One way ANOVA followed by Bonferroni test was applied to test for significance and values are expressed as mean +/- SE (p < 0.05). RESULTS: There was a significant increase in myocardial catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GPx) activities with no significant change in myocardial thiobarbituric acid reactive substances (TBARS) only in group PO 5 as compared to group C. There was no light microscopic evidence of tissue injury. A significant rise in myocardial TBARS and depletion of myocardial endogenous antioxidants (SOD, CAT and GPx) along with significant myocyte injury was observed in control rats subjected to ischemia-reperfusion (C IR). Hearts from palm olein oil fed rats subjected to ischemia-reperfusion (PO 5 IR and PO 10 IR) were protected from increase in TBARS and depletion of endogenous antioxidants as compared to C IR group. No significant myocyte injury was present in the treated groups. CONCLUSIONS: The present study demonstrated for the first time that dietary palm olein oil protected rat heart from oxidative stress associated with ischemic-reperfusion injury. [Abstract/Link to Full Text]

Mickley GA, Kenmuir CL, McMullen CA, Snyder A, Yocom AM, Likins-Fowler D, Valentine EL, Weber B, Biada JM
Long-term age-dependent behavioral changes following a single episode of fetal N-methyl-D-Aspartate (NMDA) receptor blockade.
BMC Pharmacol. 2004 Oct 28;4(1):28.
BACKGROUND: Administration of the N-methyl-D-aspartate (NMDA) antagonist ketamine during the perinatal period can produce a variety of behavioral and neuroanatomical changes. Our laboratory has reported reliable changes in learning and memory following a single dose of ketamine administered late in gestation. However, the nature of the drug-induced changes depends on the point during embryonic development when ketamine is administered. Embryonic day 18 (E18) rat fetuses pre-treated with ketamine (100 mg/kg, i.p. through the maternal circulation) and taught a conditioned taste aversion (CTA) learn and remember the CTA, whereas E19 fetuses do not. The current study sought to determine if long-term behavioral effects could be detected in animals that received ketamine or a saline control injection on either E18 or E19. Rat behavior was evaluated on two different measures: spontaneous locomotion and water maze learning. Measurements were collected during 2 periods: Juvenile test period [pre-pubertal locomotor test: Postnatal Day 11 (P11); pre-pubertal water maze test: P18] or Young-adult test period [post-pubertal locomotor test: P60; post-pubertal water maze test: P81]. RESULTS: Water maze performance of ketamine-treated rats was similar to that of controls when tested on P18. Likewise, the age of the animal at the time of ketamine/saline treatment did not influence learning of the maze. However, the young-adult water maze test (P81) revealed reliable benefits of prenatal ketamine exposure - especially during the initial re-training trial. On the first trial of the young adult test, rats treated with ketamine on E18 reached the hidden platform faster than any other group - including rats treated with ketamine on E19. Swim speeds of experimental and control rats were not significantly different. Spontaneous horizontal locomotion measured during juvenile testing indicated that ketamine-treated rats were less active than controls. However, later in development, rats treated with ketamine on E18 were more active than rats that received the drug on E19. CONCLUSION: These data suggest that both the day in fetal development when ketamine is administered and the timing of post-natal behavioral testing interact to influence behavioral outcomes. The data also indicate that the paradoxical age-dependent effects of early ketamine treatment on learning, previously described in fetuses and neonates, may also be detected later in young adult rats. [Abstract/Link to Full Text]

Garbacki N, Tits M, Angenot L, Damas J
Inhibitory effects of proanthocyanidins from Ribes nigrum leaves on carrageenin acute inflammatory reactions induced in rats.
BMC Pharmacol. 2004 Oct 21;4(1):25.
BACKGROUND: The anti-inflammatory effects of proanthocyanidins (PACs), isolated from blackcurrant (Ribes nigrum L.) leaves, were analysed using carrageenin-induced paw oedema and carrageenin-induced pleurisy in rats. RESULTS: Pretreatment of the animals with PACs (10, 30, 60 and 100 mg/kg, i.p.) reduced paw oedema induced by carrageenin in a dose and time-dependent manner. PACs also inhibited dose-dependently carrageenin-induced pleurisy in rats. They reduced (A) lung injury, (B) pleural exudate formation, (C) polymorphonuclear cell infiltration, (D) pleural exudate levels of TNF-alpha, IL-1beta and CINC-1 but did not affect IL-6 and IL-10 levels. They reduced (E) pleural exudate levels of nitrite/nitrate (NOx). In indomethacin treated rats, the volume of pleural exudate was low, its content in leukocytes and its contents in TNF-alpha, IL-1beta, IL-6 and IL-10 but not in NOx were reduced. These data suggest that the anti-inflammatory properties of PACs are achieved through a different pattern from those of indomethacin. CONCLUSION: These results suggest that the main mechanism of the anti-inflammatory effect of PACs mainly lies in an interference with the migration of the leukocytes. Moreover, PACs inhibited in vivo nitric oxide release. [Abstract/Link to Full Text]

Perrone JA, Chabla JM, Hallas BH, Horowitz JM, Torres G
Weight loss dynamics during combined fluoxetine and olanzapine treatment.
BMC Pharmacol. 2004 Oct 21;4(1):27.
BACKGROUND: Fluoxetine and olanzapine combination therapy is rapidly becoming an effective strategy for managing symptoms of treatment-resistant depression. Determining drug-drug interactions, drug metabolism and pharmacokinetics is of particular interest for revealing potential liabilities associated with drug augmentation in special patient populations. In the current studies, we chronically administered fluoxetine and olanzapine in non-stressed rats to extend our previous findings regarding body weight dynamics. RESULTS: Chronic fluoxetine (10 mg/kg) and olanzapine (5 mg/kg and 0.5 mg/kg) treatment decreased weight gain irrespective of olanzapine dosing. At the 10 mg/kg and 5 mg/kg dose, respectively, fluoxetine and olanzapine also significantly reduced food and water consumption. This pharmacodynamic event-related effect, however, was not observed at the 10 mg/kg and 0.5 mg/kg dosing paradigm suggesting differences in tolerability rates as a function of olanzapine dose. The decrease in weight gain was not associated with apparent changes in glucose metabolism as vehicle- and drug-treated rats showed undistinguishable serum glucose levels. The combination of fluoxetine and olanzapine in rats yielded drug plasma concentrations that fell within an expected therapeutic range for these drugs in psychiatric patients. CONCLUSIONS: These data suggest that fluoxetine and olanzapine treatment decreases weight gain in rats; a pharmacodynamic event-related effect that differs considerably from what is observed in the clinical condition. The possibility of mismatched models regarding body weight changes during drug augmentation therapy should be seriously considered. [Abstract/Link to Full Text]

Kinobe R, Ji Y, Nakatsu K
Peroxynitrite-mediated inactivation of heme oxygenases.
BMC Pharmacol. 2004 Oct 21;4(1):26.
BACKGROUND: Endogenous nitric oxide (NO) and carbon monoxide (CO) are generated by nitric oxide synthase and heme oxygenase, respectively. Like NO, CO has been accepted as an important cellular signaling molecule in biological systems. An up-regulation in both gene and protein expression of heme oxygenase-1 (HO-1) under oxidative/nitrosative stress has been well documented, and the protective role of HO-1 and HO-2 against oxidative damage is proposed. However, data on the direct effect of reactive oxygen/nitrogen species (ROS/RNS) on HO function is incomplete. Using gas chromatography to quantify carbon monoxide (CO) formation from heme oxidation, we investigated the effects of peroxynitrite (ONOO-) on the in vitro catalytic activity of rat spleen (HO-1) and brain (HO-2) microsomal heme oxygenases. RESULTS: Exposure to ONOO- led to concentration-dependent but reversible decreases in the activity of microsomal rat spleen and brain HO activity. Spleen HO activity was 100-fold more sensitive to ONOO--dependent inactivation compared to that of the brain, with IC50 values of 0.015 +/- 0.005 mM and 1.25 +/- 0.25 mM respectively. Inhibition of both rat spleen and brain microsomal HO activity was also observed with tetra-nitromethane, a tyrosine nitrating agent, as well as two NO donors, S-nitrosoglutathione (GSNO) and diethylamine NONOate (DEA-NONOate). However, no additive effect was found following the application of NO donors and ONOO- together. CONCLUSION: These results indicate that ONOO- may regulate HO-1 and HO-2 activities by mechanisms that involve different interactions with these proteins. It is suggested that while nitration of tyrosine residues and oxidation of sulfhydryl groups may be involved, consideration should be given to other facets of ONOO- chemistry. This inhibition of HO activity offers a mechanism for cross talk between the nitric oxide synthase and HO systems. [Abstract/Link to Full Text]

Amantea D, Bowery NG
Reduced inhibitory action of a GABAB receptor agonist on [3H]-dopamine release from rat ventral tegmental area in vitro after chronic nicotine administration.
BMC Pharmacol. 2004 Oct 20;4(1):24.
BACKGROUND: The activation of GABAB receptors in the ventral tegmental area (VTA) has been suggested to attenuate the rewarding properties of psychostimulants, including nicotine. However, the neurochemical mechanism that underlie this effect remains unknown. Since GABAB receptors modulate the release of several neurotransmitters in the mammalian brain, we have characterised the effect of the GABAB receptor agonist baclofen on the release of [3H]-dopamine ([3H]-DA) from VTA slices of naive rats and of rats pre-treated with nicotine. RESULTS: In naive rats, baclofen concentration-dependently inhibited the electrically evoked release of [3H]-DA from the isolated VTA (EC50 = 0.103 microM, 95% CI = 0.043-0.249), without affecting the basal [3H]-monoamine overflow. This effect was mediated by activation of GABAB receptors as it was blocked by the selective receptor antagonist CGP55845A. Chronic administration of nicotine (0.4 mg kg(-1), s.c., for 14 days) affected neither the basal nor the electrically evoked release of [3H]-DA from VTA slices. However, the inhibitory effect of baclofen (10 microM) on the stimulated [3H]-monoamine overflow was abolished in rats pre-treated with nicotine as compared to saline-injected controls. CONCLUSIONS: Our results demonstrate that GABAB receptor activation reduces the release of DA from the rat VTA. In addition, a reduced sensitivity of VTA GABAB receptors appears to develop after chronic exposure to nicotine. The resulting disinhibition of VTA DA neurones might therefore contribute to the sensitised dopaminergic responses observed in the rat mesocorticolimbic system following repeated administration of nicotine. [Abstract/Link to Full Text]

Yang B, Brown KK, Chen L, Carrick KM, Clifton LG, McNulty JA, Winegar DA, Strum JC, Stimpson SA, Pahel GL
Serum adiponectin as a biomarker for in vivo PPARgamma activation and PPARgamma agonist-induced efficacy on insulin sensitization/lipid lowering in rats.
BMC Pharmacol. 2004 Oct 18;4(1):23.
BACKGROUND: PPARgamma agonists ameliorate insulin resistance and dyslipidemia in type 2 diabetic patients. Adiponectin possesses insulin sensitizing properties, and predicts insulin sensitivity of both glucose and lipid metabolism. In diet-induced insulin resistant rats and ZDF rats, the current studies determined the correlation between PPARgamma agonist-upregulated fatty acid binding protein(FABP3) mRNA in adipose tissue and PPARgamma agonist-elevated serum adiponectin, and the correlation between PPARgamma agonist-elevated serum adiponectin and PPARgamma agonist-mediated efficacy in insulin sensitization and lipid lowering. RESULTS: Parallel groups of SD rats were fed a high fat/sucrose (HF) diet for 4 weeks. These rats were orally treated for the later 2 weeks with vehicle, either PPARgamma agonist GI262570 (0.2-100 mg/kg, Q.D.), or GW347845 (3 mg/kg, B.I.D). Rats on HF diet showed significant increases in postprandial serum triglycerides, free fatty acids (FFA), insulin, and area under curve (AUC) of serum insulin during an oral glucose tolerance test, but showed no change in serum glucose, adiponectin, and glucose AUC. Treatment with GI262570 dose-dependently upregulated adipose FABP3 mRNA, and increased serum adiponectin. There was a position correlation between adipose FABP3 mRNA and serum adiponectin (r = 0.7350, p < 0.01). GI262570 dose-dependently decreased the diet-induced elevations in triglycerides, FFA, insulin, and insulin AUC. Treatment with GW347845 had similar effects on serum adiponectin and the diet-induced elevations. There were negative correlations for adiponectin versus triglycerides, FFA, insulin, and insulin AUC (For GI262570, r = -0.7486, -0.4581, -0.4379, and -0.3258 respectively, all p < 0.05. For GW347845, r = -0.6370, -0.6877, -0.5512, and -0.3812 respectively, all p < 0.05). In ZDF rats treated with PPARgamma agonists pioglitazone (3-30 mg/kg, B.I.D.) or GW347845 (3 mg/kg, B.I.D.), there were also negative correlations for serum adiponectin versus glucose, triglycerides, FFA (for pioglitazone, r = -0.7005, -0.8603, and -0.9288 respectively; for GW347845, r = -0.9721, -0.8483, and -0.9453 respectively, all p < 0.01). CONCLUSIONS: This study demonstrated that (a) PPARgamma agonists improved insulin sensitivity and ameliorated dyslipidemia in HF fed rats and ZDF rats, which were correlated with serum adiponectin; (b) Serum adiponectin was positively correlated with adipose FABP3 mRNA in GI262570-treated rats. These data suggest that serum adiponectin can serve as a biomarker for both in vivo PPARgamma activation and PPARgamma agonist-induced efficacy on insulin resistance and dyslipidemia in rats. [Abstract/Link to Full Text]

Litjens NH, van Strijen E, van Gulpen C, Mattie H, van Dissel JT, Thio HB, Nibbering PH
In vitro pharmacokinetics of anti-psoriatic fumaric acid esters.
BMC Pharmacol. 2004 Oct 12;422.
BACKGROUND: Psoriasis is a chronic inflammatory skin disease that can be successfully treated with a mixture of fumaric acid esters (FAE) formulated as enteric-coated tablets for oral use. These tablets consist of dimethylfumarate (DMF) and salts of monoethylfumarate (MEF) and its main bioactive metabolite is monomethylfumarate (MMF). Little is known about the pharmacokinetics of these FAE. The aim of the present study was to investigate the hydrolysis of DMF to MMF and the stability of MMF, DMF and MEF at in vitro conditions representing different body compartments. RESULTS: DMF is hydrolyzed to MMF in an alkaline environment (pH 8), but not in an acidic environment (pH 1). In these conditions MMF and MEF remained intact during the period of analysis (6 h). Interestingly, DMF was hardly hydrolyzed to MMF in a buffer of pH 7.4, but was rapidly hydrolyzed in human serum having the same pH. Moreover, in whole blood the half-life of DMF was dramatically reduced as compared to serum. The concentrations of MMF and MEF in serum and whole blood decreased with increasing time. These data indicate that the majority of the FAE in the circulation are metabolized by one or more types of blood cells. Additional experiments with purified blood cell fractions resuspended in phosphate buffered saline (pH 7.4) revealed that at concentrations present in whole blood monocytes/lymphocytes, but not granulocytes and erythrocytes, effectively hydrolyzed DMF to MMF. Furthermore, in agreement with the data obtained with the pure components of the tablet, the enteric-coated tablet remained intact at pH 1, but rapidly dissolved at pH 8. CONCLUSION: Together, these in vitro data indicate that hydrolysis of DMF to MMF rapidly occurs at pH 8, resembling that within the small intestines, but not at pH 1 resembling the pH in the stomach. At both pHs MMF and MEF remained intact. These data explain the observation that after oral FAE intake MMF and MEF, but not DMF, can be readily detected in the circulation of human healthy volunteers and psoriasis patients. [Abstract/Link to Full Text]

Gordon EM, Myers C, Blumer J
In vitro evaluation of the potential role of sulfite radical in morphine-associated histamine release.
BMC Pharmacol. 2004 Oct 6;4(1):21.
BACKGROUND: Intravenous morphine use is associated with elevated histamine release leading to bronchoconstriction, edema and hemodynamic instability in some patients. This study evaluated the possibility that sulfite, which is present as a preservative in many morphine preparations, might contribute to histamine release in vitro. RESULTS: The human mast cell line, HMC-1, was exposed to various morphine concentrations, in the absence of sulfite, under cell culture conditions. Clinically attained concentrations of morphine (0.018microg/ml and 0.45microg/ml) did not cause increased histamine release from mast cells. There was a significant increase in histamine release when the morphine concentration was increased by 1184-fold (668microg/ml morphine). Histamine release from mast cells exposed to morphine and/or sulfite required the presence of prostaglandin H synthetase. Histamine release in experiments using sulfite-containing morphine solutions was not statistically different from that observed in morphine-only solutions. CONCLUSION: Sulfite in sulfite-containing morphine solutions, at concentrations seen clinically, is not responsible for histamine release in in vitro experiments of the human mast cell line, HMC-1. This does not preclude the fact that sulfite may lead to elevation of histamine levels in vivo. [Abstract/Link to Full Text]

Chirino YI, Hernández-Pando R, Pedraza-Chaverrí J
Peroxynitrite decomposition catalyst ameliorates renal damage and protein nitration in cisplatin-induced nephrotoxicity in rats.
BMC Pharmacol. 2004 Sep 30;4(1):20.
BACKGROUND: Oxidative stress is involved in cisplatin-nephrotoxicity. However, it has not completely established if reactive nitrogen species and nitrosative stress are involved in this experimental model. The purpose of this work was to study the role of peroxynitrite, a reactive nitrogen specie, in cisplatin-nephrotoxicity using the compound 5,10,15,20-tetrakis (4-sulfonatophenyl) porphyrinato iron (III) (FeTPPS), a soluble complex able to metabolize peroxynitrite. RESULTS: In rats treated with cisplatin (a single intraperitoneal dose of 7.5 mg/kg body weight), renal nitrosative stress was made evident by the increase in 3-nitrotyrosine on day 3. In addition, cisplatin-induced nephrotoxicity was evident by the histological damage of proximal tubular cells and by the increase in (a) serum creatinine, (b) blood urea nitrogen, and (c) urinary excretion of N-acetyl-beta-D-glucosaminidase and total protein. Cisplatin-induced nitrosative stress and nephrotoxicity were attenuated by FeTPPS-treatment (15 mg/kg body weight, intraperitoneally, every 12 hours for 3 days). CONCLUSIONS: Nitrosative stress is involved in cisplatin-induced nephrotoxicity in rats. Our data suggest that peroxynitrite is involved, at least in part, in cisplatin-induced nephrotoxicity and protein nitration. [Abstract/Link to Full Text]

Recent Articles in BMC Clinical Pharmacology

Chevassus H, Mourand I, Molinier N, Lacarelle B, Brun JF, Petit P
Assessment of single-dose benzodiazepines on insulin secretion, insulin sensitivity and glucose effectiveness in healthy volunteers: a double-blind, placebo-controlled, randomized cross-over trial [ISRCTN08745124].
BMC Clin Pharmacol. 2004 Mar 4;43.
BACKGROUND: The present study aimed at investigating in healthy volunteers the effects of diazepam and clonazepam on beta-cell function, insulin sensitivity and glucose effectiveness based on the frequently sampled intravenous (0.5 gkg-1) glucose tolerance test with minimal-model analysis. METHODS: The study was designed as a double-blind, placebo-controlled, cross-over clinical trial. Diazepam (10 mg) and clonazepam (1 mg) were infused during 30 min to 15 male subjects with a mean age of 22 years (range: 20-29), after informed consent was given. Benzodiazepines were assayed by capillary gas chromatography with electron capture, insulin by radioimmunoassay and glucose by the enzymatic glucose oxidase method. RESULTS: Both benzodiazepines induced significant psychotropic effects. The acute insulin responses (AIR) were significantly and negatively correlated with the clonazepam plasma concentrations (r = -0.609, P < 0.05, n = 14). However, the mean AIR was not significantly different between the benzodiazepine-treated subjects and the controls. In addition, the parameters of glucose assimilation were significantly decreased as compared with placebo in the subgroup of 7 subjects with plasma clonazepam concentrations higher than 6.0 ng ml-1 (median and lower limit of effective therapeutic concentrations): 1.37 +/- 0.3 versus 2.84 +/- 0.60 x 10(-2)min-1 (P = 0.028) for the coefficient of glucose tolerance (Kg), 2.18 +/- 0.29 versus 3.71 +/- 0.89 x 10(-4)microUml-1min-1 (P = 0.018) for insulin sensitivity (Si) and 1.80 +/- 0.39 versus 3.59 +/- 0.71 x 10(-2)min-1 (P = 0.028) for glucose effectiveness at basal insulin (Sg). These parameters were not significantly modified when diazepam was administered; plasma levels of this drug however, were below the effective therapeutic concentrations (300 ng ml-1) from min 15 after the end of the perfusion. CONCLUSION: The present results suggest that a benzodiazepine, in particular clonazepam, may alter insulin secretion and insulin sensitivity after a single administration in healthy volunteers. [Abstract/Link to Full Text]

Levitt DG
Physiologically based pharmacokinetic modeling of arterial - antecubital vein concentration difference.
BMC Clin Pharmacol. 2004 Feb 19;42.
BACKGROUND: Modeling of pharmacokinetic parameters and pharmacodynamic actions requires knowledge of the arterial blood concentration. In most cases, experimental measurements are only available for a peripheral vein (usually antecubital) whose concentration may differ significantly from both arterial and central vein concentration. METHODS: A physiologically based pharmacokinetic (PBPK) model for the tissues drained by the antecubital vein (referred to as "arm") is developed. It is assumed that the "arm" is composed of tissues with identical properties (partition coefficient, blood flow/gm) as the whole body tissues plus a new "tissue" representing skin arteriovenous shunts. The antecubital vein concentration depends on the following parameters: the fraction of "arm" blood flow contributed by muscle, skin, adipose, connective tissue and arteriovenous shunts, and the flow per gram of the arteriovenous shunt. The value of these parameters was investigated using simultaneous experimental measurements of arterial and antecubital concentrations for eight solutes: ethanol, thiopental, 99Tcm-diethylene triamine pentaacetate (DTPA), ketamine, D2O, acetone, methylene chloride and toluene. A new procedure is described that can be used to determine the arterial concentration for an arbitrary solute by deconvolution of the antecubital concentration. These procedures are implemented in PKQuest, a general PBPK program that is freely distributed RESULTS: One set of "standard arm" parameters provides an adequate description of the arterial/antecubital vein concentration for ethanol, DTPA, thiopental and ketamine. A significantly different set of "arm" parameters was required to describe the data for D2O, acetone, methylene chloride and toluene - probably because the "arm" is in a different physiological state. CONCLUSIONS: Using the set of "standard arm" parameters, the antecubital vein concentration can be used to determine the whole body PBPK model parameters for an arbitrary solute without any additional adjustable parameters. Also, the antecubital vein concentration can be used to estimate the arterial concentration for an arbitrary input for solutes for which no arterial concentration data is available. [Abstract/Link to Full Text]

Olsen H, Andersen A, Nordbř A, Kongsgaard UE, Břrmer OP
Pharmaceutical-grade albumin: impaired drug-binding capacity in vitro.
BMC Clin Pharmacol. 2004 Mar 29;44.
BACKGROUND: Albumin is the most abundant protein in blood plasma, and due to its ligand binding properties, serves as a circulating depot for endogenous and exogenous (e.g. drugs) compounds. Hence, the unbound drug is the pharmacologically active drug. Commercial human albumin preparations are frequently used during surgery and in critically ill patients. Recent studies have indicated that the use of pharmaceutical-grade albumin is controversial in critically ill patients. In this in vitro study we investigated the drug binding properties of pharmaceutical-grade albumins (Baxter/Immuno, Octapharma, and Pharmacia & Upjohn), native human serum, and commercially available human serum albumin from Sigma Chemical Company. METHODS: The binding properties of the various albumin solutions were tested in vitro by means of ultrafiltration. Naproxen, warfarin, and digitoxin were used as ligands. HPLC was used to quantitate the total and free drug concentrations. The data were fitted to a model of two classes of binding sites for naproxen and warfarin and one class for digitoxin, using Microsoft Excel and Graphpad Prism. RESULTS: The drugs were highly bound to albumin (95-99.5%). The highest affinity (lowest K1) was found with naproxen. Pharmaceutical-grade albumin solutions displayed significantly lower drug-binding capacity compared to native human serum and Sigma albumin. Thus, the free fraction was considerably higher, approximately 40 times for naproxen and 5 and 2 times for warfarin and digitoxin, respectively. The stabilizers caprylic acid and N-acetyl-DL-tryptophan used in the manufacturing procedure seem to be of importance. Adding the stabilizers to human serum and Sigma albumin reduced the binding affinity whereas charcoal treatment of the pharmaceutical-grade albumin from Octapharma almost restored the specific binding capacity. CONCLUSION: This in vitro study demonstrates that the specific binding for warfarin and digitoxin is significantly reduced and for naproxen no longer detectable in pharmaceutical-grade albumin. It further shows that the addition of stabilizers may be of major importance for this effect. [Abstract/Link to Full Text]

Nilsson GH, Björholt I
Occurrence and quality of anticoagulant treatment of chronic atrial fibrillation in primary health care in Sweden: a retrospective study on electronic patient records.
BMC Clin Pharmacol. 2004 Feb 9;41.
BACKGROUND: Chronic atrial fibrillation is a prevalent cardiac disorder. The literature indicates varying proportions of those treated with anticoagulants, and varying intensity of anticoagulation. Electronic patient records are providing us with clinical data concerning management of anticoagulant treatment in real-life practice that is useful for audits. We aimed to assess warfarin treatment for chronic atrial fibrillation in primary health care with regard to prevalence, incidence, the proportion treated and the quality of anticoagulation control. METHODS: Five primary health care centres in Stockholm with a registered population of 75146 participated in a one-year retrospective study of electronic patient records up until May 2000. All patients over 18 years of age with an encounter labelled 'Atrial fibrillation' were identified, and all records of patients on warfarin treatment were manually reviewed. Main outcome measures were number of patients with chronic atrial fibrillation, number of patients on wafarin treatment, and time within the therapeutic prothrombin range. RESULTS: In total, 419 patients had chronic atrial fibrillation, giving a prevalence of 0.60% (age-adjusted 0.62%), the age group 65 years or older accounted for 91.6%, and 50.1% were women. Out of these, 50.4% (211 patients) were established on warfarin treatment for chronic atrial fibrillation (0.28% of the population), and there was a predominance of men (p = 0.02). Fifty-four patients started treatment with warfarin for chronic atrial fibrillation (0.07% of the population). Among 25 randomly selected patients on established treatment, the proportion of time within the therapeutic range was 70.2%. Among 24 randomly selected patients starting treatment, the proportion of time with therapeutic values was 54.2% and 66.9% the first and second months of treatment, respectively. CONCLUSIONS: Chronic atrial fibrillation is common among the elderly in primary health care, and about half of these patients are treated with warfarin. It appears to be under-diagnosed, and may also be under-treated. About two thirds of treatment time is spent within the therapeutic range, and further improvement of the quality of anticoagulation control with warfarin may therefore be hard to achieve. [Abstract/Link to Full Text]

Mills E, Prousky J, Raskin G, Gagnier J, Rachlis B, Montori VM, Juurlink D
The safety of over-the-counter niacin. A randomized placebo-controlled trial [ISRCTN18054903].
BMC Clin Pharmacol. 2003 Nov 13;34.
BACKGROUND: Niacin is widely available over the counter (OTC). We sought to determine the safety of 500 mg immediate release niacin, when healthy individuals use them as directed. METHODS: 51 female and 17 male healthy volunteers (mean age 27 years SD 4.4) participated in a randomized placebo-controlled blinded trial of a single dose of an OTC, immediate-release niacin 500 mg (n = 33), or a single dose of placebo (n = 35) on an empty stomach. The outcomes measured were self-reported incidence of flushing and other adverse effects. RESULTS: 33 volunteers on niacin (100%) and 1 volunteer on placebo (3%) flushed (relative risk 35, 95% confidence interval (CI) 6.8-194.7). Mean time to flushing on niacin was 18.2 min (95% CI: 12.7-23.6); mean duration of flushing was 75.4 min (95% CI: 62.5-88.2). Other adverse effects occurred commonly in the niacin group: chills (51.5% vs. 0%, P <.0001), generalized pruritus (75% vs. 0%, P = <.001), gastrointestinal upset (30% vs. 3%, P =.005), and cutaneous tingling (30% vs. 0%, P = <.001). Six participants did not tolerate the adverse effects of niacin and 3 required medical attention. CONCLUSION: Clinicians counseling patients about niacin should alert patients not only about flushing but also about gastrointestinal symptoms, the most severe in this study. They should not trust that patients would receive information about these side effects or their prevention (with aspirin) from the OTC packet insert. [Abstract/Link to Full Text]

Levitt DG
The pharmacokinetics of the interstitial space in humans.
BMC Clin Pharmacol. 2003 Jul 30;33.
BACKGROUND: The pharmacokinetics of extracellular solutes is determined by the blood-tissue exchange kinetics and the volume of distribution in the interstitial space in the different organs. This information can be used to develop a general physiologically based pharmacokinetic (PBPK) model applicable to most extracellular solutes. METHODS: The human pharmacokinetic literature was surveyed to tabulate the steady state and equilibrium volume of distribution of the solutes mannitol, EDTA, morphine-6-glucuronide, morphine-3-glucuronide, inulin and beta-lactam antibiotics with a range of protein binding (amoxicillin, piperacillin, cefatrizine, ceforanide, flucloxacillin, dicloxacillin). A PBPK data set was developed for extracellular solutes based on the literature for interstitial organ volumes. The program PKQuest was used to generate the PBPK model predictions. The pharmacokinetics of the protein (albumin) bound beta-lactam antibiotics were characterized by two parameters: 1) the free fraction of the solute in plasma; 2) the interstitial albumin concentration. A new approach to estimating the capillary permeability is described, based on the pharmacokinetics of the highly protein bound antibiotics. RESULTS: About 42% of the total body water is extracellular. There is a large variation in the organ distribution of this water - varying from about 13% of total tissue water for skeletal muscle, up to 70% for skin and connective tissue. The weakly bound antibiotics have flow limited capillary-tissue exchange kinetics. The highly protein bound antibiotics have a significant capillary permeability limitation. The experimental pharmacokinetics of the 11 solutes is well described using the new PBPK data set and PKQuest. CONCLUSIONS: Only one adjustable parameter (systemic clearance) is required to completely characterize the PBPK for these extracellular solutes. Knowledge of just this systemic clearance allows one to predict the complete time course of the absolute drug concentrations in the major organs. PKQuest is freely available [Abstract/Link to Full Text]

Kumar A, Choudhuri G, Aggarwal R
Piperacillin induced bone marrow suppression: a case report.
BMC Clin Pharmacol. 2003 Jun 5;32.
BACKGROUND: Piperacillin (and piperacillin/tazobactam) is a commonly prescribed antibiotic and is generally considered safe. We report a case of piperacillin induced bone marrow suppression. CASE PRESENTATION: A 19-year-old boy was being treated with piperacillin followed by piperacillin/tazobactam for infected pancreatic pseudocyst. After 21 days of treatment, he developed neutropenia and thrombocytopenia. These reversed promptly after stopping piperacillin/tazobactam. The time course of events suggested that piperacillin was the cause of bone marrow suppression in this patient. CONCLUSION: Bone marrow suppression is a serious adverse effect of piperacillin, which should be kept in mind while treating patients with this drug. [Abstract/Link to Full Text]

Levitt DG
The use of a physiologically based pharmacokinetic model to evaluate deconvolution measurements of systemic absorption.
BMC Clin Pharmacol. 2003 Mar 19;31.
BACKGROUND: An unknown input function can be determined by deconvolution using the systemic bolus input function (r) determined using an experimental input of duration ranging from a few seconds to many minutes. The quantitative relation between the duration of the input and the accuracy of r is unknown. Although a large number of deconvolution procedures have been described, these routines are not available in a convenient software package. METHODS: Four deconvolution methods are implemented in a new, user-friendly software program (PKQuest, Three of these methods are characterized by input parameters that are adjusted by the user to provide the "best" fit. A new approach is used to determine these parameters, based on the assumption that the input can be approximated by a gamma distribution. Deconvolution methodologies are evaluated using data generated from a physiologically based pharmacokinetic model (PBPK). RESULTS AND CONCLUSIONS: The 11-compartment PBPK model is accurately described by either a 2 or 3-exponential function, depending on whether or not there is significant tissue binding. For an accurate estimate of r the first venous sample should be at or before the end of the constant infusion and a long (10 minute) constant infusion is preferable to a bolus injection. For noisy data, a gamma distribution deconvolution provides the best result if the input has the form of a gamma distribution. For other input functions, good results are obtained using deconvolution methods based on modeling the input with either a B-spline or uniform dense set of time points. [Abstract/Link to Full Text]

Niihara Y, Ge J, Shalev O, Wu H, Tu A, Tanaka KR
Desferrioxamine decreases NAD redox potential of intact red blood cells: evidence for desferrioxamine as an inducer of oxidant stress in red blood cells.
BMC Clin Pharmacol. 2002 Oct 24;28.
BACKGROUND: Desferrioxamine (DFO) is an important iron chelating agent. It has also been thought of as an agent with anti-oxidant potential as it chelates ferric iron in various parts of the body. However, there is evidence suggesting that it may paradoxically affect red blood cells (RBC) by inducing intracellular oxidant stress. To further understand the mechanism of DFO's interaction with RBC, we conducted a study to determine the effect of DFO upon RBC's redox status. METHODS: We examined NAD redox potential in intact RBC (N = 5) incubated with DFO. RBC were incubated with 6 mM DFO for 2 hours. RESULTS: Significant decreases in NAD redox potential were observed after incubation of RBC with 6 mM DFO. The mean decrease was 10.01 PlusMinus; 1.98% (p < 0.0004). CONCLUSIONS: The data confirm the oxidant effect of DFO on RBC. [Abstract/Link to Full Text]

Levitt DG
PKQuest: capillary permeability limitation and plasma protein binding - application to human inulin, dicloxacillin and ceftriaxone pharmacokinetics.
BMC Clin Pharmacol. 2002 Sep 26;27.
BACKGROUND: It is generally assumed that the tissue exchange of antibiotics is flow limited (complete equilibration between the capillary and the tissue water). This assumption may not be valid if there is a large amount of plasma protein binding because the effective capillary permeability depends on the product of the intrinsic capillary permeability (PS) and the fraction of solute that is free in the blood (fwB). PKQuest, a new generic physiologically based pharmacokinetic software routine (PBPK), provides a novel approach to modeling capillary permeability in which the only adjustable parameter is the PS of muscle. METHODS: All the results were obtained by applying PKQuest to previously published human pharmacokinetic data. RESULTS: The PKQuest analysis suggests that the highly protein bound antibiotics dicloxacillin and ceftriaxone have a significant capillary permeability limitation. The human muscle capillary PS of inulin, dicloxacillin and ceftriaxone was 0.6, 13 and 6 ml/min/100 gm, respectively. The ceftriaxone protein binding is non-linear, saturating at high plasma concentrations. The experimental ceftriaxone data over a wide range of intravenous inputs (0.15 to 3 gms) was well described by PKQuest. PKQuest is the first PBPK that includes both permeability limitation and non-linear binding. CONCLUSIONS: Because of their high degree of plasma protein binding, dicloxacillin and ceftriaxone appear to have a diffusion limited exchange rate between the blood and tissue and are not flow limited as had been previously assumed. PKQuest and all the examples are freely available at http:\\ [Abstract/Link to Full Text]

Loke YK, Derry S, Pritchard-Copley A
Appetite suppressants and valvular heart disease - a systematic review.
BMC Clin Pharmacol. 2002 Aug 23;26.
BACKGROUND: Although appetite suppressants have been implicated in the development of valvular heart disease, the exact level of risk is still uncertain. Initial studies suggested that as many as 1 in 3 exposed patients were affected, but subsequent research has yielded substantially different figures. Our objective was to systematically assess the risk of valvular heart disease with appetite suppressants. METHODS: We accepted studies involving obese patients treated with any of the following appetite suppressants: fenfluramine, dexfenfluramine, and phentermine. Three types of studies were reviewed: controlled and uncontrolled observational studies, and randomized controlled trials. Outcomes of interest were echocardiographically detectable aortic regurgitation of mild or greater severity, or mitral regurgitation of moderate or greater severity. RESULTS: Of the 1279 patients evaluated in seven uncontrolled cohort studies, 236 (18%) and 60 (5%) were found to have aortic and mitral regurgitation, respectively. Pooled data from six controlled cohort studies yielded, for aortic regurgitation, a relative risk ratio of 2.32 (95% confidence intervals 1.79 to 3.01, p < 0.00001) and an attributable rate of 4.9%, and for mitral regurgitation, a relative risk ratio of 1.55 (95% confidence intervals 1.06 to 2.25, p = 0.02) with an attributable rate of 1.0%. Only one case of valvular heart disease was detected in 57 randomized controlled trials, but this was judged unrelated to drug therapy. CONCLUSIONS: The risk of valvular heart disease is significantly increased by the appetite suppressants reviewed here. Nevertheless, when considering all the evidence, valvulopathy is much less common than suggested by the initial, less methodologically rigorous studies. [Abstract/Link to Full Text]

Levitt DG
PKQuest: measurement of intestinal absorption and first pass metabolism - application to human ethanol pharmacokinetics.
BMC Clin Pharmacol. 2002 Aug 15;24.
BACKGROUND: PKQuest, a new physiologically based pharmacokinetic (PBPK) program, is applied to human ethanol data. The classical definition of first pass metabolism (FPM) based on the differences in the area under the curve (AUC) for identical intravenous and oral doses is invalid if the metabolism is non-linear (e.g. ethanol). Uncertainties in the measurement of FPM have led to controversy about the magnitude of gastric alcohol metabolism. PKQuest implements a new, rigorous definition of FPM based on finding the equivalent intravenous input function that would produce a blood time course identical to that observed for the oral intake. This input function equals the peripheral availability (PA) and the FPM is defined by: FPM = Total oral dose - PA. PKQuest also provides a quantitative measurement of the time course of intestinal absorption. METHODS: PKQuest was applied to previously published ethanol pharmacokinetic data. RESULTS: The rate of ethanol absorption is primarily limited by the rate of gastric emptying. For oral ethanol with a meal: absorption is slow (Tilde; 3 hours) and the fractional PKQuest FPM was 36% (0.15 gm/Kg dose) and 7% (0.3 gm/Kg). In contrast, fasting oral ethanol absorption is fast (Tilde; 50 minutes) and FPM is small. CONCLUSIONS: The standard AUC and one compartment methods significantly overestimate the FPM. Gastric ethanol metabolism is not significant. Ingestion of a coincident meal with the ethanol can reduce the peak blood level by about 4 fold at low doses. PKQuest and all the examples are freely available on the web at [Abstract/Link to Full Text]

Levitt DG
PKQuest: a general physiologically based pharmacokinetic model. Introduction and application to propranolol.
BMC Clin Pharmacol. 2002 Aug 15;25.
BACKGROUND: A "physiologically based pharmacokinetic" (PBPK) approach uses a realistic model of the animal to describe the pharmacokinetics. Previous PBPKs have been designed for specific solutes, required specification of a large number of parameters and have not been designed for general use. METHODS: This new PBPK program (PKQuest) includes a "Standardhuman" and "Standardrat" data set so that the user input is minimized. It has a simple user interface, graphical output and many new features: 1) An option that uses the measured plasma concentrations to solve for the time course of the gastrointestinal, intramuscular, intraperotineal or skin absorption and systemic availability of a drug - for a general non-linear system. 2) Capillary permeability limitation defined in terms of the permeability-surface area products. 4) Saturable plasma and tissue protein binding. 5) A lung model that includes perfusion-ventilation mismatch. 6) A general optimization routine using either a global (simulated annealing) or local (Powell) minimization applicable to all model parameters. RESULTS: PKQuest was applied to measurements of human propranolol pharmacokinetics and intestinal absorption. A meal has two effects: 1) increases portal blood flow by 50%; and 2) decreases liver metabolism by 20%. There is a significant delay in the oval propranolol absorption in fasting subjects that is absent in fed subjects. The oral absorption of the long acting form of propranolol continues for a period of more than 24 hours. CONCLUSIONS: PKQuest provides a new general purpose, easy to use, freely distributed and physiologically rigorous PBPK software routine. [Abstract/Link to Full Text]

Ward A, Caro JJ, Green TC, Huybrechts K, Arana A, Wait S, Eleftheriou A
An international survey of patients with thalassemia major and their views about sustaining life-long desferrioxamine use.
BMC Clin Pharmacol. 2002 Apr 23;23.
BACKGROUND: Management of thalassemia major requires patients to have life-long access to a treatment regimen of regular blood transfusions coupled with iron chelation therapy. The objective of this study was to investigate patients' reasons for missing iron chelation therapy with desferrioxamine, and the support to sustain life-long adherence to treatment. METHODS: From October 1999 to May 2000 a survey of patients with thalassemia major was conducted in ten countries: Cyprus, Egypt, Greece, Hong Kong, India, Iran, Italy, Jordan, Taiwan, and the United States. RESULTS: 1,888 questionnaires (65%) were returned. Most patients (1,573) used desferrioxamine, and 79% administered a dose at least 4 days a week. Inaccessibility of the drug was a common reason for missing a dose in India (51%), and in Iran (25%), whereas, in any other country, it was a reason for less than 17% of patients. Overall, 58% reported reasons for missing a dose related to their beliefs or feelings about the medication, and 42% drug-related side effects. CONCLUSION: Many patients miss doses of desferrioxamine and an opportunity remains to develop interventions that provide more support to sustain use of desferrioxamine. [Abstract/Link to Full Text]

Ammon S, Marx C, Behrens C, Hofmann U, Mürdter T, Griese EU, Mikus G
Diclofenac does not interact with codeine metabolism in vivo: a study in healthy volunteers.
BMC Clin Pharmacol. 2002 Feb 27;22.
BACKGROUND: Previously, we have demonstrated a marked inhibition of codeine glucuronidation by diclofenac in human liver tissue homogenate. We therefore aimed to investigate whether diclofenac inhibits glucuronidation of codeine also in vivo in healthy volunteers. METHODS: In a randomised, placebo-controlled, double-blind, cross-over study, 12 healthy volunteers received a singe of 100 mg codeine phosphate plus 50 mg diclofenac sodium or codeine phosphate plus placebo. Over a 36 hour period serum concentrations of codeine and its metabolites as well as urinary excretion were analysed using LC-mass spectrometry. Side effects were recorded and analgesic efficacy was determined using the cold pressor test (0-6 h). RESULTS: A single dose of diclofenac did not alter the formation of codeine-6-glucuronide in healthy volunteers. Metabolic clearance of codeine to morphine was not affected by diclofenac. In terms of side effects, both treatments were well tolerated. Diclofenac did not significantly influence the analgesic effects of codeine in the cold pressor test. CONCLUSIONS: In contrast to recent in vitro data, a single oral dose of diclofenac did not alter the glucuronidation of codeine in healthy volunteers. [Abstract/Link to Full Text]

Wiwanitkit V, Wangsaturaka D, Tangphao O
LDL-cholesterol lowering effect of a generic product of simvastatin compared to simvastatin (Zocor) in Thai hypercholesterolemic subjects -- a randomized crossover study, the first report from Thailand.
BMC Clin Pharmacol. 2002;21.
BACKGROUND: It is commonly agreed that people with a high blood LDL-cholesterol will have a higher risk of coronary artery disease (CAD) than people with low blood LDL-cholesterol. Due to the increasingly high costs of medication in Thailand, the government has set up several measures to combat the problem. One of such strategies is to promote the utilization of locally manufactured drug products, especially those contained in the National Drug List. Simvastatin, an HMG-CoA reductase inhibitor, is listed as an essential drug for the treatment of hypercholesterolemia. Here, we reported the study on the LDL-cholesterol-lowering effect of a generic simvastatin product in comparison with the Zocor, in 43 healthy thai volunteers. METHOD: The generic product tested was Eucor, locally manufactured by Greater Pharma Ltd., Part, Thailand, and the reference product was Zocor (Merck Sharp & Dohme, USA). The two products were administered as 10-mg single oral doses in a two-period crossover design. After drug administration, serial blood samples were collected every 4 weeks for 16 weeks. The major parameter monitored in this study was blood LDL-cholesterol. RESULT: After taking the drugs for the first 8 weeks, no statistically significant difference was detected in blood LDL-cholesterol between the first (Zocor-treated) and the second (Eucor-treated) groups. After crossover and taking drugs for further 8 weeks, a similar result was obtained, i.e., no significant difference in blood LDL-cholesterol between the first (Eucor-treated) and the second (Zocor-treated) groups was observed. Upon completion of the 16-week study, there was also no statistically significant difference in the changes of all tested blood parameters between the two products (randomized block ANOVA, N = 37). Only minor side effects, mainly dizziness and nausea, were observed in both products. CONCLUSION: Our study demonstrated no significant differences in the therapeutic effect and safety between the generic and original simvastatin products. [Abstract/Link to Full Text]

Tagwireyi DD, Ball DE, Nhachi CF
Routine prophylactic antibiotic use in the management of snakebite.
BMC Clin Pharmacol. 2001;14.
BACKGROUND: Routine antibiotic prophylaxis following snakebite is not recommended but evidence suggests that it may be common practice in Zimbabwe. This study set out to determine and describe the extent of this practice at Parirenyatwa Hospital, a large teaching hospital in Zimbabwe METHODS: A retrospective case review (1996 to 1999 inclusive) of all cases of snakebite was undertaken at Parirenyatwa Hospital. Cases with a diagnosis of snakebite, presenting within 24 hours of the bite and with no complications or concurrent illness were defined as "routine prophylactic antibiotic use". RESULTS: From 78 cases which satisfied the inclusion criteria, 69 (88.5%) received antibiotics. Ten different antibiotics from 6 different classes were used with penicillins the most commonly prescribed (benzylpenicillin in 29% of cases, alone or in combination). Over 40% of antibiotics were given parenterally although all patients were conscious on admission. The total cost of antibiotics used was estimated at US$522.98. CONCLUSION: Routine prophylactic use of antibiotics in snakebite at Parirenyatwa Hospital is common practice. This may highlight the lack of a clearly defined policy leading to wasteful inappropriate antibiotic use which is costly and may promote bacterial antibiotic resistance. Further work is required to investigate the reasons for this practice and to design appropriate interventions to counter it. [Abstract/Link to Full Text]

Loke YK, Derry S
Reporting of adverse drug reactions in randomised controlled trials - a systematic survey.
BMC Clin Pharmacol. 2001;13.
BACKGROUND: Decisions on treatment are guided, not only by the potential for benefit, but also by the nature and severity of adverse drug reactions. However, some researchers have found numerous deficiencies in trial reports of adverse effects. We sought to confirm these findings by evaluating trials of drug therapy published in seven eminent medical journals in 1997. METHODS: Literature review to determine whether the definition, recording and reporting of adverse drug reactions in clinical trials were in accordance with published recommendations on structured reporting. RESULTS: Of the 185 trials reviewed, 25 (14%) made no mention of adverse drug reactions. Data in a further 60 (32%) could not be fully evaluated, either because numbers were not given for each treatment arm (31 trials), or because a generic statement was made without full details (29 trials). When adverse drug reactions such as clinical events or patient symptoms were mentioned in the reports, details on how they had been recorded were given in only 14/95 (15%) and 18/104 (17%) trials respectively. Of the 86 trials that mentioned severity of adverse drug reactions, only 42 (49%) stated how severity had been defined. The median amount of space used for safety data in the Results and Discussion sections was 5.8%. CONCLUSIONS: Trial reports often failed to provide details on how adverse drug reactions were defined or recorded. The absence of such methodological information makes comparative evaluation of adverse reaction rates potentially unreliable. Authors and journals should adopt recommendations on the structured reporting of adverse effects. [Abstract/Link to Full Text]

Mieczkowski T, Tsatsakis AM, Kruger M, Psillakis T
The concentration of three anti-seizure medications in hair: the effects of hair color, controlling for dose and age.
BMC Clin Pharmacol. 2001;12.
BACKGROUND: This paper assess the relationship between the quantity of three anti-seizure medications in hair and the color of the analyzed hair, while controlling for the effects of dose, dose duration, and patient age for 140 clinical patients undergoing anti-seizure therapy. Three drugs are assessed: carbamazepine (40 patients), valproic acid (40 patients), and phenytoin (60 patients). The relationship between hair assay results, hair color, dose, dose duration, and age is modeled using an analysis of covariance. The covariance model posits the hair assay results as the dependent variable, the hair color as the qualitative categorical independent variable, and dose, dose duration, and age as covariates. The null hypothesis assessed is that there is a no relationship between hair color and the quantity of analyte determined by hair assay such that darker colored hair will demonstrate higher concentrations of analyte than lighter colored hair. RESULTS: The analysis reveals that there is a significant relationship between dose and concentration for all hair color categories independent of the other covariates or the categorical independent variable. CONCLUSION: There does not appear to be any relationship between carbamazepine concentration and hair color. There is a weak relationship between hair color and valproic acid concentration, which the data suggest may be mediated by age. There is a significant, moderate relationship between phenytoin concentration and hair color such that darker colored hair has greater concentration values than lighter colored hair. [Abstract/Link to Full Text]

de Abajo FJ, García Rodríguez LA
Risk of upper gastrointestinal bleeding and perforation associated with low-dose aspirin as plain and enteric-coated formulations.
BMC Clin Pharmacol. 2001;11.
BACKGROUND: The use of low-dose aspirin has been reported to be associated with an increased risk of upper gastrointestinal complications (UGIC). The coating of aspirin has been proposed as an approach to reduce such a risk. To test this hypothesis, we carried out a population based case-control study. METHODS: We identified incident cases of UGIC (bleeding or perforation) aged 40 to 79 years between April 1993 to October 1998 registered in the General Practice Research Database. Controls were selected randomly from the source population. Adjusted estimates of relative risk (RR) associated with current use of aspirin as compared to non use were computed using unconditional logistic regression. RESULTS: We identified 2,105 cases of UGIC and selected 11,500 controls. Among them, 287 (13.6%) cases and 837 (7.3%) controls were exposed to aspirin, resulting in an adjusted RR of 2.0 (1.7-2.3). No clear dose-effect was found within the range of 75-300 mg. The RR associated with enteric-coated formulations (2.3, 1.6-3.2) was similar to the one of plain aspirin (1.9, 1.6-2.3), and no difference was observed depending on the site. The first two months of treatment was the period of greater risk (RR= 4.5, 2.9-7.1). The concomitant use of aspirin with high-dose NSAIDs greatly increased the risk of UGIC (13.3, 8.5-20.9) while no interaction was apparent with low-medium doses (2.2, 1.0-4.6). CONCLUSIONS: Low-dose aspirin increases by twofold the risk of UGIC in the general population and its coating does not modify the effect. Concomitant use of low-dose aspirin and NSAIDs at high doses put patients at a specially high risk of UGIC. [Abstract/Link to Full Text]

Recent Articles in BMC Medicine

Avasarala JR, O'Donovan CA, Roach SE, Camacho F, Feldman SR
Analysis of NAMCS data for multiple sclerosis, 1998-2004.
BMC Med. 2007;56.
BACKGROUND: To our knowledge, no study to date has investigated the prescribing patterns of immunomodulatory agents (IMAs) in an outpatient setting in the United States. To address this issue, we performed retrospective data analyses on National Ambulatory Medical Care Survey (NAMCS) data for MS patient visits between 1998 and 2004. METHODS: NAMCS data are a weighted estimate of the nationwide frequency of patients' outpatient clinic visits. We analyzed NAMCS data in the following categories: (1) the proportion of MS patient visits to neurologists, family practitioners or internists, (2) age/gender/race/geographical distribution patterns in patient visits, and (3) the proportion of patients on IMA treatment among established MS patients. RESULTS: There were an estimated 6.7 million multiple sclerosis (MS) patient visits to the clinics between 1998-2004. Neurologists recorded the most patient visits, 50.7%. Patient visits were mostly in the fourth and fifth decade age group (57.9%). The male to female ratio was 1:4. No statistical evidence was observed for a decline or increase in IMA usage. About 62% patients visiting neurologists and 92% seen by family practitioners/internists were not using IMAs. Our results suggest that between the years 1998-2003, the use of interferon-1a tended to decline while the use of interferon-1b and glatiramer acetate, increased. CONCLUSION: Strategies that lead to improved use of IMAs in the management of MS in the outpatient setting are needed. [Abstract/Link to Full Text]

Sermet-Gaudelus I, Renouil M, Fajac A, Bidou L, Parbaille B, Pierrot S, Davy N, Bismuth E, Reinert P, Lenoir G, Lesure JF, Rousset JP, Edelman A
In vitro prediction of stop-codon suppression by intravenous gentamicin in patients with cystic fibrosis: a pilot study.
BMC Med. 2007;55.
BACKGROUND: Cystic fibrosis (CF) is caused by mutations in the gene encoding the cystic fibrosis transmembrane conductance regulator (CFTR) protein, which acts as a chloride channel activated by cyclic AMP (cAMP). The most frequent mutation found in 70% of CF patients is F508del, while premature stop mutations are found in about 10% of patients. In vitro aminoglycoside antibiotics (e.g. gentamicin) suppress nonsense mutations located in CFTR permitting translation to continue to the natural termination codon. Pharmacologic suppression of stop mutations within the CFTR may be of benefit to a significant number of patients. Our pilot study was conducted to determine whether intravenous gentamicin suppresses stop codons in CF patients and whether it has clinical benefits. METHODS: A dual gene reporter system was used to determine the gentamicin-induced readthrough level of the most frequent stop mutations within the CFTR in the French population. We investigated readthrough efficiency in response to 10 mg/kg once-daily intravenous gentamicin perfusions in patients with and without stop mutations. Respiratory function, sweat chloride concentration, nasal potential difference (NPD) and CFTR expression in nasal epithelial cells were measured at baseline and after 15 days of treatment. RESULTS: After in vitro gentamicin incubation, the readthrough efficiency for the Y122X mutation was at least five times higher than that for G542X, R1162X, and W1282X. In six of the nine patients with the Y122X mutation, CFTR immunodetection showed protein at the membrane of the nasal epithelial cells and the CFTR-dependent Cl- secretion in NPD measurements increased significantly. Respiratory status also improved in these patients, irrespective of the gentamicin sensitivity of the bacteria present in the sputum. Mean sweat chloride concentration decreased significantly and normalised in two patients. Clinical status, NPD and sweat Cl- values did not change in the Y122X patients with no protein expression, in patients with the other stop mutations investigated in vitro and those without stop mutations. CONCLUSION: Suppression of stop mutations in the CFTR gene with parenteral gentamicin can be predicted in vitro and is associated with clinical benefit and significant modification of the CFTR-mediated Cl- transport in nasal and sweat gland epithelium. [Abstract/Link to Full Text]

Norton ML, Saltman DC
Corrections in an electronic environment.
BMC Med. 2007;54.
The need to publish corrections to scientific articles, and occasionally to retract them, has been recognized for decades. However very little emphasis has been attached to how this is done, provided that the retraction or correction is accessible. We are considering a policy to directly correct our online publications. [Abstract/Link to Full Text]

Meissner K, Distel H, Mitzdorf U
Evidence for placebo effects on physical but not on biochemical outcome parameters: a review of clinical trials.
BMC Med. 2007;53.
BACKGROUND: Recent reviews on placebo effects in clinical trials suggest that objective changes following placebo treatments may not exist or, at least, have been considerably overestimated. However, the possibility that yet unidentified subsets of parameters are responsive to placebo treatments has not been taken into account. Therefore, the aim of the present study is to examine the effects of placebo treatments on objectively measured outcome parameters by specifically focusing on peripheral disease processes. METHODS: An initial dataset was collected from a MEDLINE search for placebo-controlled, randomized clinical trials. Trials with stable disease conditions were identified, and the effects of placebo treatments on peripheral outcome parameters were estimated by the changes from baseline in the placebo groups. An explorative data analysis was conducted in order to identify parameter classes with differential responsiveness to placebo treatments. A subgroup meta-analysis of a second dataset was performed to test whether the preliminary classification would also apply to placebo effects derived from the comparison of placebo groups with untreated control groups. RESULTS: The explorative analysis of outcome parameters and strength of placebo effects yielded a classification into responsive "physical" versus non-responsive "biochemical" parameters. In total, 50% of trials measuring physical parameters showed significant placebo effects, compared with 6% of trials measuring biochemical parameters. A subgroup meta-analysis substantiated the differential response (physical parameters: n = 14, Hedges' pooled effect size g = 0.34, 95% CI 0.22 to 0.46; biochemical parameters: n = 15, g = 0.03, 95% CI -0.04 to 0.10). The subanalysis of the second dataset supported the classification and revealed a significant improvement for physical parameters (n = 20, g = 0.22, 95% CI 0.07 to 0.36) and a deterioration for biochemical parameters (n = 6, g = -0.17, 95% CI -0.31 to -0.02). CONCLUSION: The results suggest that placebo interventions can improve physical disease processes of peripheral organs more easily and effectively than biochemical processes. This differential response offers a good starting point for theoretical considerations on possible mediating mechanisms, and for future investigations in this field. [Abstract/Link to Full Text]

Kjaer P, Bendix T, Sorensen JS, Korsholm L, Leboeuf-Yde C
Are MRI-defined fat infiltrations in the multifidus muscles associated with low back pain?
BMC Med. 2007;52.
BACKGROUND: Because training of the lumbar muscles is a commonly recommended intervention in low back pain (LBP), it is important to clarify whether lumbar muscle atrophy is related to LBP. Fat infiltration seems to be a late stage of muscular degeneration, and can be measured in a non-invasive manner using magnetic resonance imaging. The purpose of this study was to investigate if fat infiltration in the lumbar multifidus muscles (LMM) is associated with LBP in adults and adolescents. METHODS: In total, 412 adults (40-year-olds) and 442 adolescents (13-year-olds) from the general Danish population participated in this cross-sectional cohort study. People with LBP were identified through questionnaires. Using MRI, fat infiltration of the LMM was visually graded as none, slight or severe. Odds ratios were calculated for both age groups, taking into account sex, body composition and leisure time physical activity for both groups, and physical workload (in adults only) or daily bicycling (in adolescents only). RESULTS: Fat infiltration was noted in 81% of the adults but only 14% of the adolescents. In the adults, severe fat infiltration was strongly associated with ever having had LBP (OR 9.2; 95% CI 2.0-43.2), and with having LBP in the past year (OR 4.1; 1.5-11.2), but there was no such association in adolescents. None of the investigated moderating factors had an obvious effect on the OR in the adults. CONCLUSION: Fat infiltration in the LMM is strongly associated with LBP in adults only. However, it will be necessary to quantify these measurements objectively and to investigate the direction of this link longitudinally in order to determine if the abnormal muscle is the cause of LBP or vice versa. [Abstract/Link to Full Text]

Bartels NK, Börgel J, Wieczorek S, Büchner N, Hanefeld C, Bulut D, Mügge A, Rump LC, Sanner BM, Epplen JT
Risk factors and myocardial infarction in patients with obstructive sleep apnea: impact of beta2-adrenergic receptor polymorphisms.
BMC Med. 2007;51.
BACKGROUND: The increased sympathetic nervous activity in patients with obstructive sleep apnea (OSA) is largely responsible for the high prevalence of arterial hypertension, and it is suggested to adversely affect triglyceride and high-density lipoprotein (HDL) cholesterol levels in these patients. The functionally relevant polymorphisms of the beta2-adrenergic receptor (Arg-47Cys/Arg16Gly and Gln27Glu) have been shown to exert modifying effects on these risk factors in previous studies, but results are inconsistent. METHODS: We investigated a group of 429 patients (55 +/- 10.7 years; 361 men, 68 women) with moderate to severe obstructive sleep apnea (apnea/hypopnea index (AHI) 29.1 +/- 23.1/h) and, on average, a high cardiovascular risk profile (body mass index 31.1 +/- 5.6, with hypertension in 60.1%, dyslipidemia in 49.2%, and diabetes in 17.2% of patients). We typed the beta2-adrenergic receptor polymorphisms and investigated the five most frequent haplotypes for their modifying effects on OSA-induced changes in blood pressure, heart rate, and lipid levels. The prevalence of cardiovascular risk factors and coronary heart disease (n = 55, 12.8%) and survived myocardial infarction (n = 27, 6.3%) were compared between the genotypes and haplotypes. RESULTS: Multivariate linear/logistic regressions revealed a significant and independent (from BMI, age, sex, presence of diabetes, use of antidiabetic, lipid-lowering, and antihypertensive medication) influence of AHI on daytime systolic and diastolic blood pressure, heart rate, prevalence of hypertension, and triglyceride and HDL levels. The beta2-adrenergic receptor genotypes and haplotypes showed no modifying effects on these relationships or on the prevalence of dyslipidemia, diabetes, and coronary heart disease, yet, for all three polymorphisms, heterozygous carriers had a significantly lower relative risk for myocardial infarction (Arg-47Cys: n = 195, odds ratio (OR) = 0.32, P = 0.012; Arg16Gly: n = 197, OR = 0.39, P = 0.031; Gln27Glu: OR = 0.37, P = 0.023). Carriers of the most frequent haplotype (n = 113) (haplotype 1; heterozygous for all three polymorphisms) showed a five-fold lower prevalence of survived myocardial infarction (OR = 0.21, P = 0.023). CONCLUSION: Our study showed no significant modifying effect of the functionally relevant beta2-adrenergic receptor polymorphisms on OSA-induced blood pressure, heart rate, or lipid changes. Nevertheless, heterozygosity of these polymorphisms is associated with a lower prevalence of survived myocardial infarction in this group with, on average, a high cardiovascular risk profile. [Abstract/Link to Full Text]

Provenzano PP, Eliceiri KW, Campbell JM, Inman DR, White JG, Keely PJ
Collagen reorganization at the tumor-stromal interface facilitates local invasion.
BMC Med. 2006;4(1):38.
BACKGROUND: Stromal-epithelial interactions are of particular significance in breast tissue as misregulation of these interactions can promote tumorigenesis and invasion. Moreover, collagen-dense breast tissue increases the risk of breast carcinoma, although the relationship between collagen density and tumorigenesis is not well understood. As little is known about epithelial-stromal interactions in vivo, it is necessary to visualize the stroma surrounding normal epithelium and mammary tumors in intact tissues to better understand how matrix organization, density, and composition affect tumor formation and progression. METHODS: Epithelial-stromal interactions in normal mammary glands, mammary tumors, and tumor explants in three-dimensional culture were studied with histology, electron microscopy, and nonlinear optical imaging methodologies. Imaging of the tumor-stromal interface in live tumor tissue ex vivo was performed with multiphoton laser-scanning microscopy (MPLSM) to generate multiphoton excitation (MPE) of endogenous fluorophores and second harmonic generation (SHG) to image stromal collagen. RESULTS: We used both laser-scanning multiphoton and second harmonic generation microscopy to determine the organization of specific collagen structures around ducts and tumors in intact, unfixed and unsectioned mammary glands. Local alterations in collagen density were clearly seen, allowing us to obtain three-dimensional information regarding the organization of the mammary stroma, such as radiating collagen fibers that could not have been obtained using classical histological techniques. Moreover, we observed and defined three tumor-associated collagen signatures (TACS) that provide novel markers to locate and characterize tumors. In particular, local cell invasion was found predominantly to be oriented along certain aligned collagen fibers, suggesting that radial alignment of collagen fibers relative to tumors facilitates invasion. Consistent with this observation, primary tumor explants cultured in a randomly organized collagen matrix realigned the collagen fibers, allowing individual tumor cells to migrate out along radially aligned fibers. CONCLUSION: The presentation of these tumor-associated collagen signatures allowed us to identify pre-palpable tumors and see cells at the tumor-stromal boundary invading into the stroma along radially aligned collagen fibers. As such, TACS should provide indications that a tumor is, or could become, invasive, and may serve as part of a strategy to help identify and characterize breast tumors in animal and human tissues. [Abstract/Link to Full Text]

Jabba SV, Oelke A, Singh R, Maganti RJ, Fleming S, Wall SM, Everett LA, Green ED, Wangemann P
Macrophage invasion contributes to degeneration of stria vascularis in Pendred syndrome mouse model.
BMC Med. 2006;437.
BACKGROUND: Pendred syndrome, an autosomal-recessive disorder characterized by deafness and goiter, is caused by a mutation of SLC26A4, which codes for the anion exchanger pendrin. We investigated the relationship between pendrin expression and deafness using mice that have (Slc26a4+/+ or Slc26a4+/-) or lack (Slc26a4-/-) a complete Slc26a4 gene. Previously, we reported that stria vascularis of adult Slc26a4-/- mice is hyperpigmented and that marginal cells appear disorganized. Here we determine the time course of hyperpigmentation and marginal cell disorganization, and test the hypothesis that inflammation contributes to this tissue degeneration. METHODS: Slc26a4-/- and age-matched control (Slc26a4+/+ or Slc26a4+/-) mice were studied at four postnatal (P) developmental stages: before and after the age that marks the onset of hearing (P10 and P15, respectively), after weaning (P28-41) and adult (P74-170). Degeneration and hyperpigmentation stria vascularis was evaluated by confocal microscopy. Gene expression in stria vascularis was analyzed by microarray and quantitative RT-PCR. In addition, the expression of a select group of genes was quantified in spiral ligament, spleen and liver to evaluate whether expression changes seen in stria vascularis are specific for stria vascularis or systemic in nature. RESULTS: Degeneration of stria vascularis defined as hyperpigmentation and marginal cells disorganization was not seen at P10 or P15, but occurred after weaning and was associated with staining for CD68, a marker for macrophages. Marginal cells in Slc26a4-/-, however, had a larger apical surface area at P10 and P15. No difference in the expression of Lyzs, C3 and Cd45 was found in stria vascularis of P15 Slc26a4+/- and Slc26a4-/- mice. However, differences in expression were found after weaning and in adult mice. No difference in the expression of markers for acute inflammation, including Il1a, Il6, Il12a, Nos2 and Nos3 were found at P15, after weaning or in adults. The expression of macrophage markers including Ptprc (= Cd45), Cd68, Cd83, Lyzs, Lgals3 (= Mac2 antigen), Msr2, Cathepsins B, S, and K (Ctsb, Ctss, Ctsk) and complement components C1r, C3 and C4 was significantly increased in stria vascularis of adult Slc26a4-/- mice compared to Slc26a4+/+ mice. Expression of macrophage markers Cd45 and Cd84 and complement components C1r and C3 was increased in stria vascularis but not in spiral ligament, liver or spleen of Slc26a4-/- compared to Slc26a4+/- mice. The expression of Lyzs was increased in stria vascularis and spiral ligament but not in liver or spleen. CONCLUSION: The data demonstrate that hyperpigmentation of stria vascularis and marginal cell reorganization in Slc26a4-/- mice occur after weaning, coinciding with an invasion of macrophages. The data suggest that macrophage invasion contributes to tissue degeneration in stria vascularis, and that macrophage invasion is restricted to stria vascularis and is not systemic in nature. The delayed onset of degeneration of stria vascularis suggests that a window of opportunity exists to restore/preserve hearing in mice and therefore possibly in humans suffering from Pendred syndrome. [Abstract/Link to Full Text]

Enaw JO, Zhu H, Yang W, Lu W, Shaw GM, Lammer EJ, Finnell RH
CHKA and PCYT1A gene polymorphisms, choline intake and spina bifida risk in a California population.
BMC Med. 2006;436.
BACKGROUND: Neural tube defects (NTDs) are among the most common of all human congenital defects. Over the last two decades, accumulating evidence has made it clear that periconceptional intake of folic acid can significantly reduce the risk of NTD affected pregnancies. This beneficial effect may be related to the ability of folates to donate methyl groups for critical physiological reactions. Choline is an essential nutrient and it is also a methyl donor critical for the maintenance of cell membrane integrity and methyl metabolism. Perturbations in choline metabolism in vitro have been shown to induce NTDs in mouse embryos. METHODS: This study investigated whether single nucleotide polymorphisms (SNPs) in human choline kinase A (CHKA) gene and CTP:phosphocholine cytidylytransferase (PCYT1A) gene were risk factors for spina bifida. Fluorescence-based allelic discrimination analysis was performed for the two CHKA intronic SNPs hCV1562388 (rs7928739) and hCV1562393, and PCYT1A SNP rs939883 and rs3772109. The study population consisted of 103 infants with spina bifida and 338 non-malformed control infants who were born in selected California counties in the period 1989-1991. RESULTS: The CHKA SNP hCV1562388 genotypes with at least one C allele were associated with a reduced risk of spina bifida (odds ratio = 0.60, 95%CI = 0.38-0.94). The PCYT1A SNP rs939883 genotype AA was associated with a twofold increased risk of spina bifida (odds ratio = 1.89, 95% CI = 0.97-3.67). These gene-only effects were not substantially modified by analytic consideration to maternal periconceptional choline intake. CONCLUSION: Our analyses showed genotype effects of CHKA and PCYT1A genes on spina bifida risk, but did not show evidence of gene-nutrient interactions. The underlying mechanisms are yet to be resolved. [Abstract/Link to Full Text]

Heller RF, Gemmell I, Edwards R, Buchan I, Awasthi S, Volmink JA
Prioritising between direct observation of therapy and case-finding interventions for tuberculosis: use of population impact measures.
BMC Med. 2006;435.
BACKGROUND: Population impact measures (PIMs) have been developed as tools to help policy-makers with locally relevant decisions over health risks and benefits. This involves estimating and prioritizing potential benefits of interventions in specific populations. Using tuberculosis (TB) in India as an example, we examined the population impact of two interventions: direct observation of therapy and increasing case-finding. METHODS: PIMs were calculated using published literature and national data for India, and applied to a notional population of 100,000 people. Data included the incidence or prevalence of smear-positive TB and the relative risk reduction from increasing case finding and the use of direct observation of therapy (applied to the baseline risks over the next year), and the incremental proportion of the population eligible for the proposed interventions. RESULTS: In a population of 100,000 people in India, the directly observed component of the Directly Observed Treatment, Short-course (DOTS) programme may prevent 0.188 deaths from TB in the next year compared with 1.79 deaths by increasing TB case finding. The costs of direct observation are (in international dollars) 5960 I dollars and of case finding are 4839 I dollars or 31702 I dollars and 2703 I dollars per life saved respectively. CONCLUSION: Increasing case-finding for TB will save nearly 10 times more lives than will the use of the directly observed component of DOTS in India, at a smaller cost per life saved. The demonstration of the population impact, using simple and explicit numbers, may be of value to policy-makers as they prioritize interventions for their populations. [Abstract/Link to Full Text]

Melzer D, Murray A, Hurst AJ, Weedon MN, Bandinelli S, Corsi AM, Ferrucci L, Paolisso G, Guralnik JM, Frayling TM
Effects of the diabetes linked TCF7L2 polymorphism in a representative older population.
BMC Med. 2006;434.
BACKGROUND: A polymorphism in the transcription factor 7-like 2 (TCF7L2) gene has been found to be associated with type 2 diabetes in case-control studies. We aimed to estimate associations of the marker rs7903146 (C/T) polymorphism with fasting glucose, lipids, diabetes prevalence and complications in an older general population. METHODS: In total, 944 subjects aged > or = 65 years from the population representative InCHIANTI study were enrolled in this study. Those with fasting blood glucose of > or = 7 mmol/l or physician diagnosis were considered diabetic. Cut-off points for impaired fasting glucose (IFG) were > or = 5.6 mmol/l to < 7 mmol/l. RESULTS: In the general population sample, minor (T) allele carriers of rs7903146 had higher fasting blood glucose (FBG) (p = 0.028) but lower fasting insulin (p = 0.030) and HOMA2b scores (p = 0.001), suggesting poorer beta-cell function. T allele carriers also had smaller waist circumference (p = 0.009), lower triglyceride levels (p = 0.006), and higher high-density lipoprotein cholesterol (p = 0.008).The prevalence of diabetes or IFG was 32.4% in TT carriers and 23.3% in CC carriers; adjusted OR = 1.67 (95% confidence interval 1.05 to 2.65, p = 0.031). Within the diabetic and IFG groups, fewer T allele carriers had metabolic syndrome features (p = 0.047) or had experienced a myocardial infarction (p = 0.037). Conversely, T allele carriers with diabetes had poorer renal function (reduced 24-hour creatinine clearance, p = 0.013), and possibly more retinopathy (p = 0.067). Physician-diagnosed dementia was more common in the T carriers (in diabetes p = 0.05, with IFG p = 0.024). CONCLUSION: The TCF7L2 rs7903146 polymorphism is associated with lower insulin levels, smaller waist circumference, and lower risk lipid profiles in the general elderly population. Patients with diabetes who are carriers of the minor allele are less likely to have metabolic-syndrome features, but may experience more microvascular complications, although the number of cases was small. If replicated, these findings may have implications for developing treatment approaches tailored by genotype. [Abstract/Link to Full Text]

Muscatello DJ, Searles A, MacDonald R, Jorm L
Communicating population health statistics through graphs: a randomised controlled trial of graph design interventions.
BMC Med. 2006;433.
BACKGROUND: Australian epidemiologists have recognised that lay readers have difficulty understanding statistical graphs in reports on population health. This study aimed to provide evidence for graph design improvements that increase comprehension by non-experts. METHODS: This was a double-blind, randomised, controlled trial of graph-design interventions, conducted as a postal survey. Control and intervention participants were randomly selected from telephone directories of health system employees. Eligible participants were on duty at the listed location during the study period. Controls received a booklet of 12 graphs from original publications, and intervention participants received a booklet of the same graphs with design modifications. A questionnaire with 39 interpretation tasks was included with the booklet. Interventions were assessed using the ratio of the prevalence of correct responses given by the intervention group to those given by the control group for each task. RESULTS: The response rate from 543 eligible participants (261 intervention and 282 control) was 67%. The prevalence of correct answers in the control group ranged from 13% for a task requiring knowledge of an acronym to 97% for a task identifying the largest category in a pie chart. Interventions producing the greatest improvement in comprehension were: changing a pie chart to a bar graph (3.6-fold increase in correct point reading), changing the y axis of a graph so that the upward direction represented an increase (2.9-fold increase in correct judgement of trend direction), a footnote to explain an acronym (2.5-fold increase in knowledge of the acronym), and matching the y axis range of two adjacent graphs (two-fold increase in correct comparison of the relative difference in prevalence between two population subgroups). CONCLUSION: Profound population health messages can be lost through use of overly technical language and unfamiliar statistical measures. In our study, most participants did not understand age standardisation and confidence intervals. Inventive approaches are required to address this problem. [Abstract/Link to Full Text]

Camitz M, Liljeros F
The effect of travel restrictions on the spread of a moderately contagious disease.
BMC Med. 2006;432.
BACKGROUND: Much research in epidemiology has been focused on evaluating conventional methods of control strategies in the event of an epidemic or pandemic. Travel restrictions are often suggested as an efficient way to reduce the spread of a contagious disease that threatens public health, but few papers have studied in depth the effects of travel restrictions. In this study, we investigated what effect different levels of travel restrictions might have on the speed and geographical spread of an outbreak of a disease similar to severe acute respiratory syndrome (SARS). METHODS: We used a stochastic simulation model incorporating survey data of travel patterns between municipalities in Sweden collected over 3 years. We tested scenarios of travel restrictions in which travel over distances >50 km and 20 km would be banned, taking into account different levels of compliance. RESULTS: We found that a ban on journeys >50 km would drastically reduce the speed and geographical spread of outbreaks, even when compliance is < 100%. The result was found to be robust for different rates of intermunicipality transmission intensities. CONCLUSION: This study supports travel restrictions as an effective way to mitigate the effect of a future disease outbreak. [Abstract/Link to Full Text]

Dandona L, Lakshmi V, Sudha T, Kumar GA, Dandona R
A population-based study of human immunodeficiency virus in south India reveals major differences from sentinel surveillance-based estimates.
BMC Med. 2006;431.
BACKGROUND: The human immunodeficiency virus (HIV) burden among adults in India is estimated officially by direct extrapolation of annual sentinel surveillance data from public-sector antenatal and sexually transmitted infection (STI) clinics and some high-risk groups. The validity of these extrapolations has not been systematically examined with a large sample population-based study. METHODS: We sampled 13838 people, 15-49 years old, from 66 rural and urban clusters using a stratified random method to represent adults in Guntur district in the south Indian state of Andhra Pradesh. We interviewed the sampled participants and obtained dried blood spots from them, and tested blood for HIV antibody, antigen and nucleic acid. We calculated the number of people with HIV in Guntur district based on these data, compared it with the estimate using the sentinel surveillance data and method, and analysed health services use data to understand the differences. RESULTS: In total, 12617 people (91.2% of the sampled group) gave a blood sample. Adjusted HIV prevalence was 1.72% (95% confidence interval 1.35-2.09%); men 1.74% (1.27-2.21%), women 1.70% (1.36-2.04%); rural 1.64% (1.10-2.18%), urban 1.89% (1.39-2.39%). HIV prevalence was 2.58% and 1.20% in people in the lower and upper halves of a standard of living index (SLI). Of women who had become pregnant during the past 2 years, 21.1% had used antenatal care in large public-sector hospitals participating in sentinel surveillance. There was an over-representation of the lowest SLI quartile (44.7%) in this group, and 3.61% HIV prevalence versus 1.08% in the remaining pregnant women. HIV prevalence was higher in that group even when women were matched for the same SLI half (lower half 4.39%, upper 2.63%) than in the latter (lower 1.06%, upper 1.05%), due to referral of HIV-positive/suspected women by private practitioners to public hospitals. The sentinel surveillance method (HIV prevalence: antenatal clinic 3%, STI clinic 22.8%, female sex workers 12.8%) led to an estimate of 112635 (4.38%) people with HIV, 15-49 years old, in Guntur district, which was 2.5 times the 45942 (1.79%) estimate based on our population-based study. CONCLUSION: The official method in India leads to a gross overestimation of the HIV burden in this district due to addition of substantial extra HIV estimates from STI clinics, the common practice of referral of HIV-positive/suspected people to public hospitals, and a preferential use of public hospitals by people in lower socioeconomic strata. India may be overestimating its HIV burden with the currently used official estimation method. [Abstract/Link to Full Text]

Lenz O, Fornoni A
Chronic kidney disease care delivered by US family medicine and internal medicine trainees: results from an online survey.
BMC Med. 2006;430.
BACKGROUND: Complications of chronic kidney disease (CKD) contribute to morbidity and mortality. Consequently, treatment guidelines have been developed to facilitate early detection and treatment. However, given the high prevalence of CKD, many patients with early CKD are seen by non-nephrologists, who need to be aware of CKD complications, screening methods and treatment goals in order to initiate timely therapy and referral. METHODS: We performed a web-based survey to assess perceptions and practice patterns in CKD care among 376 family medicine and internal medicine trainees in the United States. Questions were focused on the identification of CKD risk factors, screening for CKD and associated co-morbidities, as well as management of anemia and secondary hyperparathyroidism in patients with CKD. RESULTS: Our data show that CKD risk factors are not universally recognized, screening for CKD complications is not generally taken into consideration, and that the management of anemia and secondary hyperparathyroidism poses major diagnostic and therapeutic difficulties for trainees. CONCLUSION: Educational efforts are needed to raise awareness of clinical practice guidelines and recommendations for patients with CKD among future practitioners. [Abstract/Link to Full Text]

Job DE, Whalley HC, McIntosh AM, Owens DG, Johnstone EC, Lawrie SM
Grey matter changes can improve the prediction of schizophrenia in subjects at high risk.
BMC Med. 2006;429.
BACKGROUND: We hypothesised that subjects at familial high risk of developing schizophrenia would have a reduction over time in grey matter, particularly in the temporal lobes, and that this reduction may predict schizophrenia better than clinical measurements. METHODS: We analysed magnetic resonance images of 65 high-risk subjects from the Edinburgh High Risk Study sample who had two scans a mean of 1.52 years apart. Eight of these 65 subjects went on to develop schizophrenia an average of 2.3 years after their first scan. RESULTS: Changes over time in the inferior temporal gyrus gave a 60% positive predictive value (likelihood ratio >10) of developing schizophrenia compared to the overall 13% risk in the cohort as a whole. CONCLUSION: Changes in grey matter could be used as part of a predictive test for schizophrenia in people at enhanced risk for familial reasons, particularly for positive predictive power, in combination with other clinical and cognitive predictive measures, several of which are strong negative predictors. However, because of the limited number of subjects, this test requires independent replication to confirm its validity. [Abstract/Link to Full Text]

Pajonk F, Riedisser A, Henke M, McBride WH, Fiebich B
The effects of tea extracts on proinflammatory signaling.
BMC Med. 2006;428.
BACKGROUND: Skin toxicity is a common side effect of radiotherapy for solid tumors. Its management can cause treatment gaps and thus can impair cancer treatment. At present, in many countries no standard recommendation for treatment of skin during radiotherapy exists. In this study, we explored the effect of topically-applied tea extracts on the duration of radiation-induced skin toxicity. We investigated the underlying molecular mechanisms and compared effects of tea extracts with the effects of epigallocatechin-gallate, the proposed most-active moiety of green tea. METHODS: Data from 60 patients with cancer of the head and neck or pelvic region topically treated with green or black tea extracts were analyzed retrospectively. Tea extracts were compared for their ability to modulate IL-1beta, IL-6, IL-8, TNFalpha and PGE2 release from human monocytes. Effects of tea extracts on 26S proteasome function were assessed. NF-kappaB activity was monitored by EMSAs. Viability and radiation response of macrophages after exposure to tea extracts was measured by MTT assays. RESULTS: Tea extracts supported the restitution of skin integrity. Tea extracts inhibited proteasome function and suppressed cytokine release. NF-kappaB activity was altered by tea extracts in a complex, caspase-dependent manner, which differed from the effects of epigallocatechin-gallate. Additionally, both tea extracts, as well as epigallocatechin-gallate, slightly protected macrophages from ionizing radiation CONCLUSION: Tea extracts are an efficient, broadly available treatment option for patients suffering from acute radiation-induced skin toxicity. The molecular mechanisms underlying the beneficial effects are complex, and most likely not exclusively dependent on effects of tea polyphenols such as epigallocatechin-gallate. [Abstract/Link to Full Text]

Schumm WR
Neurologic adverse events associated with smallpox vaccination in the United States--response and comment on reporting of headaches as adverse events after smallpox vaccination among military and civilian personnel.
BMC Med. 2006;427.
BACKGROUND: Accurate reporting of adverse events occurring after vaccination is an important component of determining risk-benefit ratios for vaccinations. Controversy has developed over alleged underreporting of adverse events within U.S. military samples. This report examines the accuracy of adverse event rates recently published for headaches, and examines the issue of underreporting of headaches as a function of civilian or military sources and as a function of passive versus active surveillance. METHODS: A report by Sejvar et al was examined closely for accuracy with respect to the reporting of neurologic adverse events associated with smallpox vaccination in the United States. Rates for headaches were reported by several scholarly sources, in addition to Sejvar et al, permitting a comparison of reporting rates as a function of source and type of surveillance. RESULTS: Several major errors or omissions were identified in Sejvar et al. The count of civilian subjects vaccinated and the totals of both civilians and military personnel vaccinated were reported incorrectly by Sejvar et al. Counts of headaches reported in VAERS were lower (n = 95) for Sejvar et al than for Casey et al (n = 111) even though the former allegedly used 665,000 subjects while the latter used fewer than 40,000 subjects, with both using approximately the same civilian sources. Consequently, rates of nearly 20 neurologic adverse events reported by Sejvar et al were also incorrectly calculated. Underreporting of headaches after smallpox vaccination appears to increase for military samples and for passive adverse event reporting systems. CONCLUSION: Until revised or corrected, the rates of neurologic adverse events after smallpox vaccinated reported by Sejvar et al must be deemed invalid. The concept of determining overall rates of adverse events by combining small civilian samples with large military samples appears to be invalid. Reports of headaches as adverse events after smallpox vaccination appear to be have occurred much less frequently using passive surveillance systems and by members of the U.S. military compared to civilians, especially those employed in healthcare occupations. Such concerns impact risk-benefit ratios associated with vaccines and weigh against making vaccinations mandatory, without informed consent, even among military members. Because of the issues raised here, adverse event rates derived solely or primarily from U.S. Department of Defense reporting systems, especially passive surveillance systems, should not be used, given better alternatives, for making public health policy decisions. [Abstract/Link to Full Text]

Carrat F, Luong J, Lao H, Sallé AV, Lajaunie C, Wackernagel H
A 'small-world-like' model for comparing interventions aimed at preventing and controlling influenza pandemics.
BMC Med. 2006;426.
BACKGROUND: With an influenza pandemic seemingly imminent, we constructed a model simulating the spread of influenza within the community, in order to test the impact of various interventions. METHODS: The model includes an individual level, in which the risk of influenza virus infection and the dynamics of viral shedding are simulated according to age, treatment, and vaccination status; and a community level, in which meetings between individuals are simulated on randomly generated graphs. We used data on real pandemics to calibrate some parameters of the model. The reference scenario assumes no vaccination, no use of antiviral drugs, and no preexisting herd immunity. We explored the impact of interventions such as vaccination, treatment/prophylaxis with neuraminidase inhibitors, quarantine, and closure of schools or workplaces. RESULTS: In the reference scenario, 57% of realizations lead to an explosive outbreak, lasting a mean of 82 days (standard deviation (SD) 12 days) and affecting 46.8% of the population on average. Interventions aimed at reducing the number of meetings, combined with measures reducing individual transmissibility, would be partly effective: coverage of 70% of affected households, with treatment of the index patient, prophylaxis of household contacts, and confinement to home of all household members, would reduce the probability of an outbreak by 52%, and the remaining outbreaks would be limited to 17% of the population (range 0.8%-25%). Reactive vaccination of 70% of the susceptible population would significantly reduce the frequency, size, and mean duration of outbreaks, but the benefit would depend markedly on the interval between identification of the first case and the beginning of mass vaccination. The epidemic would affect 4% of the population if vaccination started immediately, 17% if there was a 14-day delay, and 36% if there was a 28-day delay. Closing schools when the number of infections in the community exceeded 50 would be very effective, limiting the size of outbreaks to 10% of the population (range 0.9%-22%). CONCLUSION: This flexible tool can help to determine the interventions most likely to contain an influenza pandemic. These results support the stockpiling of antiviral drugs and accelerated vaccine development. [Abstract/Link to Full Text]

Bogaty P, Brophy JM
Acute ischemic heart disease and interventional cardiology: a time for pause.
BMC Med. 2006;425.
BACKGROUND: A major change has occurred in the last few years in the therapeutic approach to patients presenting with all forms of acute coronary syndromes. Whether or not these patients present initially to tertiary cardiac care centers, they are now routinely referred for early coronary angiography and increasingly undergo percutaneous revascularization. This practice is driven primarily by the angiographic image and technical feasibility. Concomitantly, there has been a decline in expectant or ischemia-guided medical management based on specific clinical presentation, response to initial treatment, and results of noninvasive stratification. This 'tertiarization' of acute coronary care has been fueled by the increasing sophistication of the cardiac armamentarium, the peer-reviewed publication of clinical studies purporting to show the superiority of invasive cardiac interventions, and predominantly supporting (non-peer-reviewed) editorials, newsletters, and opinion pieces. DISCUSSION: This review presents another perspective, based on a critical reexamination of the evidence. The topics addressed are: reperfusion treatment of ST-elevation myocardial infarction; the indications for invasive intervention following thrombolysis; the role of invasive management in non-ST-elevation myocardial infarction and unstable angina; and cost-effectiveness and real world considerations. A few cases encountered in recent practice in community and tertiary hospitals are presented for illustrative purposes The numerous and far-reaching scientific, economic, and philosophical implications that are a consequence of this marked change in clinical practice as well as healthcare, decisional and conflict of interest issues are explored. SUMMARY: The weight of evidence does not support the contemporary unfocused broad use of invasive interventional procedures across the spectrum of acute coronary clinical presentations. Excessive and unselective recourse to these procedures has deleterious implications for the organization of cardiac health care and undesirable economic, scientific and intellectual consequences. It is suggested that there is need for a new equilibrium based on more refined clinical risk stratification in the treatment of patients who present with acute coronary syndromes. [Abstract/Link to Full Text]

van Geel AC, Geusens PP, Nagtzaam IF, Schreurs CM, van der Voort DJ, Rinkens PE, Kester AD, Dinant GJ
Timing and risk factors for clinical fractures among postmenopausal women: a 5-year prospective study.
BMC Med. 2006;424.
BACKGROUND: Many risk factors for fractures have been documented, including low bone-mineral density (BMD) and a history of fractures. However, little is known about the short-term absolute risk (AR) of fractures and the timing of clinical fractures. Therefore, we assessed the risk and timing of incident clinical fractures, expressed as 5-year AR, in postmenopausal women. METHODS: In total, 10 general practice centres participated in this population-based prospective study. Five years after a baseline assessment, which included clinical risk factor evaluation and BMD measurement, 759 postmenopausal women aged between 50 and 80 years, were re-examined, including undergoing an evaluation of clinical fractures after menopause. Risk factors for incident fractures at baseline that were significant in univariate analyses were included in a multivariate Cox survival regression analysis. The significant determinants were used to construct algorithms. RESULTS: In the total group, 12.5% (95% confidence interval (CI) 10.1-14.9) of the women experienced a new clinical fracture. A previous clinical fracture after menopause and a low BMD (T-score <-1.0) were retained as significant predictors with significant interaction. Women with a recent previous fracture (during the past 5 years) had an AR of 50.1% (95% CI 42.0-58.1) versus 21.2% (95% CI 20.7-21.6) if the previous fracture had occurred earlier. In women without a fracture history, the AR was 13.8% (95% CI 10.9-16.6) if BMD was low and 7.0% (95% CI 5.5-8.5) if BMD was normal. CONCLUSION: In postmenopausal women, clinical fractures cluster in time. One in two women with a recent clinical fracture had a new clinical fracture within 5 years, regardless of BMD. The 5-year AR for a first clinical fracture was much lower and depended on BMD. [Abstract/Link to Full Text]

Neild GH, Rodriguez-Justo M, Wall C, Connolly JO
Hyper-IgG4 disease: report and characterisation of a new disease.
BMC Med. 2006;423.
BACKGROUND: We highlight a chronic inflammatory disease we call 'hyper-IgG4 disease', which has many synonyms depending on the organ involved, the country of origin and the year of the report. It is characterized histologically by a lymphoplasmacytic inflammation with IgG4-positive cells and exuberant fibrosis, which leaves dense fibrosis on resolution. A typical example is idiopathic retroperitoneal fibrosis, but the initial report in 2001 was of sclerosing pancreatitis. METHODS: We report an index case with fever and severe systemic disease. We have also reviewed the histology of 11 further patients with idiopathic retroperitoneal fibrosis for evidence of IgG4-expressing plasma cells, and examined a wide range of other inflammatory conditions and fibrotic diseases as organ-specific controls. We have reviewed the published literature for disease associations with idiopathic, systemic fibrosing conditions and the synonyms: pseudotumour, myofibroblastic tumour, plasma cell granuloma, systemic fibrosis, xanthofibrogranulomatosis, and multifocal fibrosclerosis. RESULTS: Histology from all 12 patients showed, to varying degrees, fibrosis, intense inflammatory cell infiltration with lymphocytes, plasma cells, scattered neutrophils, and sometimes eosinophilic aggregates, with venulitis and obliterative arteritis. The majority of lymphocytes were T cells that expressed CD8 and CD4, with scattered B-cell-rich small lymphoid follicles. In all cases, there was a significant increase in IgG4-positive plasma cells compared with controls. In two cases, biopsies before and after steroid treatment were available, and only scattered plasma cells were seen after treatment, none of them expressing IgG4. Review of the literature shows that although pathology commonly appears confined to one organ, patients can have systemic symptoms and fever. In the active period, there is an acute phase response with a high serum concentration of IgG, and during this phase, there is a rapid clinical response to glucocorticoid steroid treatment. CONCLUSION: We believe that hyper-IgG4 disease is an important condition to recognise, as the diagnosis can be readily verified and the outcome with treatment is very good. [Abstract/Link to Full Text]

Hernández-Díaz S, García Rodríguez LA
Cardioprotective aspirin users and their excess risk of upper gastrointestinal complications.
BMC Med. 2006;422.
BACKGROUND: To balance the cardiovascular benefits from low-dose aspirin against the gastrointestinal harm caused, studies have considered the coronary heart disease risk for each individual but not their gastrointestinal risk profile. We characterized the gastrointestinal risk profile of low-dose aspirin users in real clinical practice, and estimated the excess risk of upper gastrointestinal complications attributable to aspirin among patients with different gastrointestinal risk profiles. METHODS: To characterize aspirin users in terms of major gastrointestinal risk factors (i.e., advanced age, male sex, prior ulcer history and use of non-steroidal anti-inflammatory drugs), we used The General Practice Research Database in the United Kingdom and the Base de Datos para la Investigación Farmacoepidemiológica en Atención Primaria in Spain. To estimate the baseline risk of upper gastrointestinal complications according to major gastrointestinal risk factors and the excess risk attributable to aspirin within levels of these factors, we used previously published meta-analyses on both absolute and relative risks of upper gastrointestinal complications. RESULTS: Over 60% of aspirin users are above 60 years of age, 4 to 6% have a recent history of peptic ulcers and over 13% use other non-steroidal anti-inflammatory drugs. The estimated average excess risk of upper gastrointestinal complications attributable to aspirin is around 5 extra cases per 1,000 aspirin users per year. However, the excess risk varies in parallel to the underlying gastrointestinal risk and might be above 10 extra cases per 1,000 person-years in over 10% of aspirin users. CONCLUSION: In addition to the cardiovascular risk, the underlying gastrointestinal risk factors have to be considered when balancing harms and benefits of aspirin use for an individual patient. The gastrointestinal harms may offset the cardiovascular benefits in certain groups of patients where the gastrointestinal risk is high and the cardiovascular risk is low. [Abstract/Link to Full Text]

Bleich A, Gelkopf M, Melamed Y, Solomon Z
Mental health and resiliency following 44 months of terrorism: a survey of an Israeli national representative sample.
BMC Med. 2006;421.
BACKGROUND: Israeli citizens have been exposed to intense and ongoing terrorism since September 2000. We previously studied the mental health impact of terrorism on the Israeli population (Bleich et al., 2002), however the long-term impact of ongoing terrorism has not yet been examined. The present study evaluated the psychological sequelae of 44 months of terrorism in Israel, and sought to identify factors that may contribute to vulnerability and resilience. METHODS: This was a telephone survey using strata sampling of 828 households, which reached a representative sample of 702 adult Israeli residents (84.8% contact rate). In total, 501 people (60.5%) agreed to participate. The methodology was similar to that of our previous study. Exposure to terrorism and other traumatic events, number of traumatic stress-related symptoms (TSRS), percentage of respondents with symptom criteria for post-traumatic stress disorder (PTSD), traumatic stress (TS) resiliency and feelings of depression, anxiety, optimism, sense of safety, and help-seeking were the main outcome measures. RESULTS: In total, 56 participants (11.2%) were directly exposed to a terrorist incident, and 101 (20.2%) had family members or friends exposed. Respondents reported a mean +/- SD of 5.0 +/- 4.5 TSRS; 45 (9%) met symptom criteria for PTSD; and 72 (14.4%) were TS-resilient. There were 147 participants (29.5%) who felt depressed, 50 (10.4%) felt anxious, and almost half (235; 47%) felt life-threatening danger; 48 (9.7%) felt the need for professional help. Women and people of Arab ethnicity had more TSRS, more PTSD, and less TS resiliency. Injury following a life-threatening experience, a major stressful life event, and a major loss of income were associated with PTSD. Immigrant status, lower education, low sense of safety, low sense of social support, high societal distress, and injury following life-threatening experiences were associated with TSRS. TSRS did not increase with exposure severity. This study revealed less depression and functional impairment, similar rates of PTSD, increased help-seeking and poorer TSRS and TS resiliency than our initial study, 2 years previously. DISCUSSION: The response of people in Israel to 4 years of terrorism is heterogeneous. Vulnerability factors change over time; Arab ethnicity, immigrant status and less education, not found to be risk factors in our previous study, were found in the present study to contribute to trauma-related distress. Prior experience of highly stressful events increases vulnerability to adverse psychological effects of terror. [Abstract/Link to Full Text]

Risberg G, Hamberg K, Johansson EE
Gender perspective in medicine: a vital part of medical scientific rationality. A useful model for comprehending structures and hierarchies within medical science.
BMC Med. 2006;420.
BACKGROUND: During the past few decades, research has reported gender bias in various areas of clinical and academic medicine. To prevent such bias, a gender perspective in medicine has been requested, but difficulties and resistance have been reported from implementation attempts. Our study aimed at analysing this resistance in relation to what is considered good medical research. METHOD: We used a theoretical model, based on scientific competition, to understand the structures of scientific medicine and how they might influence the resistance to a gender perspective in medicine. The model was originally introduced to discuss how pluralism improves rationality in the social sciences. RESULTS: The model provided a way to conceptualise different fields of research in medicine: basic research, applied research, medical philosophy, and 'empowering' research. It clarified how various research approaches within medicine relate to each other, and how they differ and compete. It also indicated why there might be conflicts between them: basic and applied research performed within the biomedical framework have higher status than gender research and other research approaches that are performed within divergent research paradigms. CONCLUSION: This hierarchy within medical research contributes to the resistance to a gender perspective, causing gender bias and making medical scientific rationality suboptimal. We recommend that the theoretical model can be applied in a wider medical context when different and hierarchically arranged research traditions are in conflict. In this way, the model might contribute to shape a medical community where scientific pluralism is acknowledged to enlarge, not to disturb, the scientific rationality of medicine. [Abstract/Link to Full Text]

Hensing G, Andersson L, Brage S
Increase in sickness absence with psychiatric diagnosis in Norway: a general population-based epidemiologic study of age, gender and regional distribution.
BMC Med. 2006;419.
BACKGROUND: The aim of this study was to assess the incidence of sickness absence with psychiatric diagnoses from 1994-2000, and the distribution across gender, age groups, diagnostic groups and regions in a general population. METHODS: The population at risk was defined as all individuals aged 16-66 years who were entitled to sickness benefits in 1994, 1996, 1998 and 2000 (n = 2,282,761 in 2000). All individuals with a full-time disability pension were excluded. The study included approximately 77% of the Norwegian population aged 16-66 years. For each year, the study base started on 1 January and ended on 31 December. Individuals that were sick-listed for more than 14/16 consecutive days with a psychiatric diagnosis on their medical certificate were selected as cases. Included in this study were data for Norway, the capital city Oslo and five regions in the southeast of the country. RESULTS: Sickness absence with psychiatric diagnoses increased in all age groups, in women and men, and in all regions. At the national level, the cumulative incidence increased in women from 1.7% in 1994 to 4.6% in 2000, and in men from 0.8% in 1994 to 2.2% in 2000. The highest cumulative incidence was found in middle-aged women and men (30-59 years). Women had a higher incidence than men in all stratification groups. The cumulative incidences in 2000 varied between 4.6% to 5.6% in women in the different regions, and for men the corresponding figures were 2.1% to 3.2%. Throughout the four years studied, women in Oslo had more than twice as high incidence levels of sickness absence with alcohol and drug diagnoses as the country as a whole. There were some differences between regions in sickness absence with specific psychiatric diagnoses, but they were small and most comparisons were non-significant. CONCLUSION: Sickness absence with psychiatric diagnoses increased between 1994 and 2000 in Norway. The increase was highest in the middle-aged, and in women. Few regional differences were found. That the increase pervaded all stratification groups supports general explanations of the increase, such as changes in attitudes to psychiatric disorders in both patients and doctors, and increased mental distress probably associated with societal changes at a more structural level. [Abstract/Link to Full Text]

Virji A, Yarnall KS, Krause KM, Pollak KI, Scannell MA, Gradison M, Řstbye T
Use of email in a family practice setting: opportunities and challenges in patient- and physician-initiated communication.
BMC Med. 2006;418.
BACKGROUND: Electronic mail (email) has the potential to improve communication between physicians and patients. METHODS: We conducted two research studies in a family practice setting: 1) a brief, anonymous patient survey of a convenience sample to determine the number of clinic patients receptive to communicating with their physician via email, and 2) a randomized, controlled pilot study to assess the feasibility of providing health education via email to family practice patients. RESULTS: Sixty-eight percent of patients used email, and the majority of those (80%) were interested in using email to communicate with the clinic. The majority also reported that their email address changed less frequently than their home address (65%, n = 173) or telephone number (68%, n = 181). Forty-two percent were willing to pay an out-of-pocket fee to have email access to their physicians. When evaluating email initiated by the clinic, 26% of otherwise eligible patients could not participate because they lacked email access; those people were more likely to be black and to be insured through Medicaid. Twenty-four subjects agreed to participate, but one-third failed to return the required consent form by mail. All participants who received the intervention emails said they would like to receive health education emails in the future. CONCLUSION: Our survey results show that patients are interested in email communication with the family practice clinic. Our feasibility study also illustrates important challenges in physician-initiated electronic communication. The 'digital divide' - decreased access to electronic technologies in lower income groups - is an ethical concern in the use of email for patient-physician communication. [Abstract/Link to Full Text]

Michiels B, Philips H, Coenen S, Yane F, Steinhauser T, Stuyck S, Denekens J, Van Royen P
The effect of giving influenza vaccination to general practitioners: a controlled trial [NCT00221676].
BMC Med. 2006;417.
BACKGROUND: No efficacy studies of influenza vaccination given to GPs have yet been published. Therefore, our purpose was to assess the effect of an inactivated influenza vaccine given to GPs on the rate of clinical respiratory tract infections (RTIs) and proven influenza cases (influenza positive nose and throat swabs and a 4-fold titre rise), while adjusting for important covariates. METHODS: In a controlled trial during two consecutive winter periods (2002-2003 and 2003-2004) we compared (77 and 100) vaccinated with (45 and 40) unvaccinated GPs working in Flanders, Belgium. Influenza antibodies were measured immediately prior to and 3-5 weeks after vaccination, as well as after the influenza epidemic. During the influenza epidemic, GPs had to record their contact with influenza cases and their own RTI symptoms every day. If they became ill, the GPs had to take nose and throat swabs during the first 4 days. We performed a multivariate regression analysis for covariates using Generalized Estimating Equations. RESULTS: One half of the GPs (vaccinated or not) developed an RTI during the 2 influenza epidemics. During the two influenza periods, 8.6% of the vaccinated and 14.7% of the unvaccinated GPs had positive swabs for influenza (RR: 0.59; 95%CI: 0.28 - 1.24). Multivariate analysis revealed that influenza vaccination prevented RTIs and swab-positive influenza only among young GPs (ORadj: 0.35; 95%CI: 0.13 - 0.96 and 0.1; 0.01 - 0.75 respectively for 30-year-old GPs). Independent of vaccination, a low basic antibody titre against influenza (ORadj 0.57; 95%CI: 0.37 - 0.89) and the presence of influenza cases in the family (ORadj 9.24; 95%CI: 2.91 - 29) were highly predictive of an episode of swab-positive influenza. CONCLUSION: Influenza vaccination was shown to protect against proven influenza among young GPs. GPs, vaccinated or not, who are very vulnerable to influenza are those who have a low basic immunity against influenza and, in particular, those who have family members who develop influenza. [Abstract/Link to Full Text]

Hall P, Ploner A, Bjöhle J, Huang F, Lin CY, Liu ET, Miller LD, Nordgren H, Pawitan Y, Shaw P, Skoog L, Smeds J, Wedrén S, Ohd J, Bergh J
Hormone-replacement therapy influences gene expression profiles and is associated with breast-cancer prognosis: a cohort study.
BMC Med. 2006;416.
BACKGROUND: Postmenopausal hormone-replacement therapy (HRT) increases breast-cancer risk. The influence of HRT on the biology of the primary tumor, however, is not well understood. METHODS: We obtained breast-cancer gene expression profiles using Affymetrix human genome U133A arrays. We examined the relationship between HRT-regulated gene profiles, tumor characteristics, and recurrence-free survival in 72 postmenopausal women. RESULTS: HRT use in patients with estrogen receptor (ER) protein positive tumors (n = 72) was associated with an altered regulation of 276 genes. Expression profiles based on these genes clustered ER-positive tumors into two molecular subclasses, one of which was associated with HRT use and had significantly better recurrence free survival despite lower ER levels. A comparison with external data suggested that gene regulation in tumors associated with HRT was negatively correlated with gene regulation induced by short-term estrogen exposure, but positively correlated with the effect of tamoxifen. CONCLUSION: Our findings suggest that post-menopausal HRT use is associated with a distinct gene expression profile related to better recurrence-free survival and lower ER protein levels. Tentatively, HRT-associated gene expression in tumors resembles the effect of tamoxifen exposure on MCF-7 cells. [Abstract/Link to Full Text]

Andrew MK, Freter SH, Rockwood K
Prevalence and outcomes of delirium in community and non-acute care settings in people without dementia: a report from the Canadian Study of Health and Aging.
BMC Med. 2006;415.
BACKGROUND: While delirium is common among older adults in acute care hospitals, its prevalence in other settings has been less well studied. We examined delirium prevalence and outcomes in a large cohort of older Canadians living outside of acute care. METHODS: In this secondary analysis of the Canadian Study of Health and Aging, the prevalence of clinically diagnosed delirium was estimated and five-year survival was compared with that of individuals with dementia of graded severity. RESULTS: Delirium was very uncommon (prevalence <0.5%) and was associated with reduced survival, similar to that of moderate-to-severe dementia. CONCLUSION: In this cohort of older Canadians, delirium in non-demented people was associated with very low 5-year survival, at levels comparable with advanced dementia. Although it is common in hospital, delirium is uncommon among older adults in their usual place of residence, suggesting that it is a potent stimulus to seek medical care. [Abstract/Link to Full Text]

Recent Articles in BMC Complementary and Alternative Medicine

Hamre HJ, Witt CM, Glockmann A, Ziegler R, Willich SN, Kiene H
Anthroposophic medical therapy in chronic disease: a four-year prospective cohort study.
BMC Complement Altern Med. 2007;710.
BACKGROUND: The short consultation length in primary care is a source of concern, and the wish for more consultation time is a common reason for patients to seek complementary medicine. Physicians practicing anthroposophic medicine have prolonged consultations with their patients, taking an extended history, addressing constitutional, psychosocial, and biographic aspect of patients' illness, and selecting optimal therapy. In Germany, health benefit programs have included the reimbursement of this additional physician time. The purpose of this study was to describe clinical outcomes in patients with chronic diseases treated by anthroposophic physicians after an initial prolonged consultation. METHODS: In conjunction with a health benefit program in Germany, 233 outpatients aged 1-74 years, treated by 72 anthroposophic physicians after a consultation of at least 30 min participated in a prospective cohort study. Main outcomes were disease severity (Disease and Symptom Scores, physicians' and patients' assessment on numerical rating scales 0-10) and quality of life (adults: SF-36, children aged 8-16: KINDL, children 1-7: KITA). Disease Score was documented after 0, 6 and 12 months, other outcomes after 0, 3, 6, 12, 18, 24, and (Symptom Score and SF-36) 48 months. RESULTS: Most common indications were mental disorders (17.6% of patients; primarily depression and fatigue), respiratory diseases (15.5%), and musculoskeletal diseases (11.6%). Median disease duration at baseline was 3.0 years (interquartile range 0.5-9.8 years). The consultation leading to study enrolment lasted 30-60 min in 51.5% (120/233) of patients and > 60 min in 48.5%. During the following year, patients had a median of 3.0 (interquartile range 1.0-7.0) prolonged consultations with their anthroposophic physicians, 86.1% (167/194) of patients used anthroposophic medication.All outcomes except KITA Daily Life subscale and KINDL showed significant improvement between baseline and all subsequent follow-ups. Improvements from baseline to 12 months were: Disease Score from mean (standard deviation) 5.95 (1.74) to 2.31 (2.29) (p < 0.001), Symptom Score from 5.74 (1.81) to 3.04 (2.16) (p < 0.001), SF-36 Physical Component Summary from 44.01 (10.92) to 47.99 (10.43) (p < 0.001), SF-36 Mental Component Summary from 42.34 (11.98) to 46.84 (10.47) (p < 0.001), and KITA Psychosoma subscale from 62.23 (19.76) to 76.44 (13.62) (p = 0.001). All these improvements were maintained until the last follow-up. Improvements were similar in patients not using diagnosis-related adjunctive therapies within the first six study months. CONCLUSION: Patients treated by anthroposophic physicians after an initial prolonged consultation had long-term reduction of chronic disease symptoms and improvement of quality of life. Although the pre-post design of the present study does not allow for conclusions about comparative effectiveness, study findings suggest that physician-provided anthroposophic therapy may play a beneficial role in the long-term care of patients with chronic diseases. [Abstract/Link to Full Text]

Mbwambo ZH, Moshi MJ, Masimba PJ, Kapingu MC, Nondo RS
Antimicrobial activity and brine shrimp toxicity of extracts of Terminalia brownii roots and stem.
BMC Complement Altern Med. 2007;79.
BACKGROUND: Ternimalia brownii Fresen (Combretaceae) is widely used in traditional medicine to treat bacterial, fungal and viral infections. There is a need to evaluate extracts of this plant in order to provide scientific proof for it's wide application in traditional medicine system. METHODS: Extraction of stem bark, wood and whole roots of T. brownii using solvents of increasing polarity, namely, Pet ether, dichloromethane, dichloromethane: methanol (1:1), methanol and aqua, respectively, afforded dry extracts. The extracts were tested for antifungal and antibacterial activity and for brine shrimp toxicity test. RESULTS: Extracts of the stem bark, wood and whole roots of T. brownii exhibited antibacterial activity against standard strains of Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, Salmonella typhi, and Bacillus anthracis and the fungi, Candida albicans and Cryptococcus neoformans. Aqueous extracts exhibited the strongest activity against both bacteria and fungi. Extracts of the roots and stem bark exhibited relatively mild cytotoxic activity against brine shrimp larvae with LC50 values ranging from 113.75-4356.76 and 36.12-1458.81 microg/ml, respectively. The stem wood extracts exhibited the highest toxicity against the shrimps (LC50 values 2.58-14.88 microg/ml), while that of cyclophosphamide, a standard anticancer drug, was 16.33 (10.60-25.15) microg/ml. CONCLUSION: These test results support traditional medicinal use of, especially, aqueous extracts for the treatment of conditions such as diarrhea, and gonorrhea. The brine shrimp results depict the general trend among plants of the genus Terminalia, which are known to contain cytotoxic compounds such as hydrolysable tannins. These results warrant follow-up through bioassay-directed isolation of the active principles. [Abstract/Link to Full Text]

White AR, Moody RC, Campbell JL
Acupressure for smoking cessation--a pilot study.
BMC Complement Altern Med. 2007;78.
BACKGROUND: Tobacco smoking is a serious risk to health: several therapies are available to assist those who wish to stop. Smokers who approach publicly funded stop-smoking clinics in the UK are currently offered nicotine replacement therapy (NRT) or bupropion, and group behaviour therapy, for which there is evidence of effectiveness. Acupuncture and acupressure are also used to help smokers, though a systematic review of the evidence of their effectiveness was inconclusive. The aim of this pilot project was to determine the feasibility of a study to test acupressure as an adjunct to one anti-smoking treatment currently offered, and to inform the design of the study. METHODS: An open randomised controlled pilot study was conducted within the six week group programme offered by the Smoking Advice Service in Plymouth, UK. All participants received the usual treatment with NRT and group behavioural therapy, and were randomised into three groups: group A with two auricular acupressure beads, group B with one bead, and group C with no additional therapy. Participants were taught to press the beads when they experienced cravings. Beads were worn in one ear for four weeks, being replaced as necessary. The main outcome measures assessed in the pilot were success at quitting (expired CO < or = 9 ppm), the dose of NRT used, and the rating of withdrawal symptoms using the Mood and Symptoms Scale. RESULTS: From 49 smokers attending four clinics, 24 volunteered to participate, 19 attended at least once after quitting, and seven remained to the final week. Participants who dropped out reported significantly fewer previous quit attempts, but no other significant differences. Participants reported stimulating the beads as expected during the initial days after quitting, but most soon reduced the frequency of stimulation. The discomfort caused by the beads was minor, and there were no significant side effects. There were technical problems with adhesiveness of the dressing. Reporting of NRT consumption was poor, with much missing data, but reporting of ratings of withdrawal symptom scores was nearly complete. However, these showed no significant changes or differences between groups for any week. CONCLUSION: Any effects of acupressure on smoking withdrawal, as an adjunct to the use of NRT and behavioural intervention, are unlikely to be detectable by the methods used here and further preliminary studies are required before the hypothesis can be tested. [Abstract/Link to Full Text]

Haidvogl M, Riley DS, Heger M, Brien S, Jong M, Fischer M, Lewith GT, Jansen G, Thurneysen AE
Homeopathic and conventional treatment for acute respiratory and ear complaints: a comparative study on outcome in the primary care setting.
BMC Complement Altern Med. 2007;77.
BACKGROUND: The aim of this study was to assess the effectiveness of homeopathy compared to conventional treatment in acute respiratory and ear complaints in a primary care setting. METHODS: The study was designed as an international, multi-centre, comparative cohort study of non-randomised design. Patients, presenting themselves with at least one chief complaint: acute (< or = 7 days) runny nose, sore throat, ear pain, sinus pain or cough, were recruited at 57 primary care practices in Austria (8), Germany (8), the Netherlands (7), Russia (6), Spain (6), Ukraine (4), United Kingdom (10) and the USA (8) and given either homeopathic or conventional treatment. Therapy outcome was measured by using the response rate, defined as the proportion of patients experiencing 'complete recovery' or 'major improvement' in each treatment group. The primary outcome criterion was the response rate after 14 days of therapy. RESULTS: Data of 1,577 patients were evaluated in the full analysis set of which 857 received homeopathic (H) and 720 conventional (C) treatment. The majority of patients in both groups reported their outcome after 14 days of treatment as complete recovery or major improvement (H: 86.9%; C: 86.0%; p = 0.0003 for non-inferiority testing). In the per-protocol set (H: 576 and C: 540 patients) similar results were obtained (H: 87.7%; C: 86.9%; p = 0.0019). Further subgroup analysis of the full analysis set showed no differences of response rates after 14 days in children (H: 88.5%; C: 84.5%) and adults (H: 85.6%; C: 86.6%). The unadjusted odds ratio (OR) of the primary outcome criterion was 1.40 (0.89-2.22) in children and 0.92 (0.63-1.34) in adults. Adjustments for demographic differences at baseline did not significantly alter the OR. The response rates after 7 and 28 days also showed no significant differences between both treatment groups. However, onset of improvement within the first 7 days after treatment was significantly faster upon homeopathic treatment both in children (p = 0.0488) and adults (p = 0.0001). Adverse drug reactions occurred more frequently in adults of the conventional group than in the homeopathic group (C: 7.6%; H: 3.1%, p = 0.0032), whereas in children the occurrence of adverse drug reactions was not significantly different (H: 2.0%; C: 2.4%, p = 0.7838). CONCLUSION: In primary care, homeopathic treatment for acute respiratory and ear complaints was not inferior to conventional treatment. [Abstract/Link to Full Text]

Borud EK, Alraek T, White A, Fonnebo V, Grimsgaard S
The effect of TCM acupuncture on hot flushes among menopausal women (ACUFLASH) study: a study protocol of an ongoing multi-centre randomised controlled clinical trial.
BMC Complement Altern Med. 2007;76.
BACKGROUND: After menopause, 10-20% of all women have nearly intolerable hot flushes. Long term use of hormone replacement therapy involves a health risk, and many women seek alternative strategies to relieve climacteric complaints. Acupuncture is one of the most frequently used complementary therapies in Norway. We designed a study to evaluate whether Traditional Chinese Medicine acupuncture-care together with self-care is more effective than self-care alone to relieve climacteric complaints. METHODS/DESIGN: The study is a multi-centre pragmatic randomised controlled trial with two parallel arms. Participants are postmenopausal women who document > or =7 flushes/24 hours and who are not using hormone replacement therapy or other medication that may influence flushes. According to power calculations 200 women are needed to detect a 50% reduction in flushes, and altogether 286 women will be recruited to allow for a 30% dropout rate.The treatment group receives 10 sessions of Traditional Chinese Medicine acupuncture-care and self-care; the control group will engage in self-care only. A team of experienced Traditional Chinese Medicine acupuncturists give acupuncture treatments. DISCUSSION: The study tests acupuncture as a complete treatment package including the therapeutic relationship and expectation. The intervention period lasts for 12 weeks, with follow up at 6 and 12 months. Primary endpoint is change in daily hot flush frequency in the two groups from baseline to 12 weeks; secondary endpoint is health related quality of life, assessed by the Women's Health Questionnaire. We also collect data on Traditional Chinese Medicine diagnoses, and we examine treatment experiences using a qualitative approach. Finally we measure biological variables, to examine potential mechanisms for the effect of acupuncture. The study is funded by The Research Council of Norway. [Abstract/Link to Full Text]

Kemper KJ, Dirkse D, Eadie D, Pennington M
What do clinicians want? Interest in integrative health services at a North Carolina academic medical center.
BMC Complement Altern Med. 2007;75.
BACKGROUND: Use of complementary medicine is common, consumer driven and usually outpatient focused. We wished to determine interest among the medical staff at a North Carolina academic medical center in integrating diverse therapies and services into comprehensive care. METHODS: We conducted a cross sectional on-line survey of physicians, nurse practitioners and physician assistants at a tertiary care medical center in 2006. The survey contained questions on referrals and recommendations in the past year and interest in therapies or services if they were to be provided at the medical center in the future. RESULTS: Responses were received from 173 clinicians in 26 different departments, programs and centers. There was strong interest in offering several specific therapies: therapeutic exercise (77%), expert consultation about herbs and dietary supplements (69%), and massage (66%); there was even stronger interest in offering comprehensive treatment programs such as multidisciplinary pain management (84%), comprehensive nutritional assessment and advice (84%), obesity/healthy lifestyle promotion (80%), fit for life (exercise and lifestyle program, 76%), diabetes healthy lifestyle promotion (73%); and comprehensive psychological services for stress management, including hypnosis and biofeedback (73%). CONCLUSION: There is strong interest among medical staff at an academic health center in comprehensive, integrated services for pain, obesity, and diabetes and in specific services in fitness, nutrition and stress management. Future studies will need to assess the cost-effectiveness of such services, as well as their financial sustainability and impact on patient satisfaction, health and quality of life. [Abstract/Link to Full Text]

Clement YN, Morton-Gittens J, Basdeo L, Blades A, Francis MJ, Gomes N, Janjua M, Singh A
Perceived efficacy of herbal remedies by users accessing primary healthcare in Trinidad.
BMC Complement Altern Med. 2007;74.
BACKGROUND: The increasing global popularity of herbal remedies requires further investigation to determine the probable factors driving this burgeoning phenomenon. We propose that the users' perception of efficacy is an important factor and assessed the perceived efficacy of herbal remedies by users accessing primary health facilities throughout Trinidad. Additionally, we determined how these users rated herbal remedies compared to conventional allopathic medicines as being less, equally or more efficacious. METHODS: A descriptive cross-sectional study was undertaken at 16 randomly selected primary healthcare facilities throughout Trinidad during June-August 2005. A de novo, pilot-tested questionnaire was interviewer-administered to confirmed herbal users (previous or current). Stepwise multiple regression analysis was done to determine the influence of predictor variables on perceived efficacy and comparative efficacy with conventional medicines. RESULTS: 265 herbal users entered the study and cited over 100 herbs for the promotion of health/wellness and the management of specific health concerns. Garlic was the most popular herb (in 48.3% of the sample) and was used for the common cold, cough, fever, as 'blood cleansers' and carminatives. It was also used in 20% of hypertension patients. 230 users (86.8%) indicated that herbs were efficacious and perceived that they had equal or greater efficacy than conventional allopathic medicines. Gender, ethnicity, income and years of formal education did not influence patients' perception of herb efficacy; however, age did (p = 0.036). Concomitant use of herbs and allopathic medicines was relatively high at 30%; and most users did not inform their attending physician. CONCLUSION: Most users perceived that herbs were efficacious, and in some instances, more efficacious than conventional medicines. We suggest that this perception may be a major contributing factor influencing the sustained and increasing popularity of herbs. Evidence-based research in the form of randomized controlled clinical trials should direct the proper use of herbs to validate (or otherwise) efficacy and determine safety. In the Caribbean, most indigenous herbs are not well investigated and this points to the urgent need for biomedical investigations to assess the safety profile and efficacy of our popular medicinal herbs. [Abstract/Link to Full Text]

Ong VY, Tan BK
Novel phytoandrogens and lipidic augmenters from Eucommia ulmoides.
BMC Complement Altern Med. 2007;73.
BACKGROUND: Plants containing compounds such as the isoflavonoids, with female hormone-like effects that bind to human estrogen receptors, are known. But none has been previously shown to have corresponding male hormone-like effects that interact with the human androgen receptor. Here, we report that the tree bark (cortex) of the Gutta-Percha tree Eucommia ulmoides possesses bimodal phytoandrogenic and hormone potentiating effects by lipidic components. METHODS: The extracts of E. ulmoides were tested using in-vitro reporter gene bioassays and in-vivo animal studies. Key compounds responsible for the steroidogenic effects were isolated and identified using solid phase extraction (SPE), high performance liquid chromatography (HPLC), thin layer chromatography (TLC), gas chromatography-mass spectroscopy (GC-MS), electron spray ionisation-mass spectroscopy (ESI-MS) and nuclear magnetic resonance (NMR). RESULTS: The following bioactivities of E. ulmoides were found: (1) a phenomenal tripartite synergism exists between the sex steroid receptors (androgen and estrogen receptors), their cognate steroidal ligands and lipidic augmenters isolated from E. ulmoides, (2) phytoandrogenic activity of E. ulmoides was mediated by plant triterpenoids binding cognately to the androgen receptor (AR) ligand binding domain. CONCLUSION: In addition to well-known phytoestrogens, the existence of phytoandrogens is reported in this study. Furthermore, a form of tripartite synergism between sex steroid receptors, sex hormones and plant-derived lipids is described for the first time. This could have contrasting clinical applications for hypogonadal- and hyperlipidaemic-related disorders. [Abstract/Link to Full Text]

Davis JM, Fleming MF, Bonus KA, Baker TB
A pilot study on mindfulness based stress reduction for smokers.
BMC Complement Altern Med. 2007;72.
BACKGROUND: Mindfulness means paying attention in the present moment, non-judgmentally, without commentary or decision-making. We report results of a pilot study designed to test the feasibility of using Mindfulness Based Stress Reduction (MBSR) (with minor modifications) as a smoking intervention. METHODS: MBSR instructors provided instructions in mindfulness in eight weekly group sessions. Subjects attempted smoking cessation during week seven without pharmacotherapy. Smoking abstinence was tested six weeks after the smoking quit day with carbon monoxide breath test and 7-day smoking calendars. Questionnaires were administered to evaluate changes in stress and affective distress. RESULTS: 18 subjects enrolled in the intervention with an average smoking history of 19.9 cigarettes per day for 26.4 years. At the 6-week post-quit visit, 10 of 18 subjects (56%) achieved biologically confirmed 7-day point-prevalent smoking abstinence. Compliance with meditation was positively associated with smoking abstinence and decreases in stress and affective distress. DISCUSSIONS AND CONCLUSION: The results of this study suggest that mindfulness training may show promise for smoking cessation and warrants additional study in a larger comparative trial. [Abstract/Link to Full Text]

Conforti A, Bellavite P, Bertani S, Chiarotti F, Menniti-Ippolito F, Raschetti R
Rat models of acute inflammation: a randomized controlled study on the effects of homeopathic remedies.
BMC Complement Altern Med. 2007;71.
BACKGROUND: One of the cardinal principles of homeopathy is the "law of similarities", according to which patients can be treated by administering substances which, when tested in healthy subjects, cause symptoms that are similar to those presented by the patients themselves. Over the last few years, there has been an increase in the number of pre-clinical (in vitro and animal) studies aimed at evaluating the pharmacological activity or efficacy of some homeopathic remedies under potentially reproducible conditions. However, in addition to some contradictory results, these studies have also highlighted a series of methodological difficulties.The present study was designed to explore the possibility to test in a controlled way the effects of homeopathic remedies on two known experimental models of acute inflammation in the rat. To this aim, the study considered six different remedies indicated by homeopathic practice for this type of symptom in two experimental edema models (carrageenan- and autologous blood-induced edema), using two treatment administration routes (sub-plantar injection and oral administration). METHODS: In a first phase, the different remedies were tested in the four experimental conditions, following a single-blind (measurement) procedure. In a second phase, some of the remedies (in the same and in different dilutions) were tested by oral administration in the carrageenan-induced edema, under double-blind (treatment administration and measurement) and fully randomized conditions. Seven-hundred-twenty male Sprague Dawley rats weighing 170-180 g were used. Six homeopathic remedies (Arnica montana D4, Apis mellifica D4, D30, Atropa belladonna D4, Hamamelis virginiana D4, Lachesis D6, D30, Phosphorus D6, D30), saline and indomethacin were tested. Edema was measured using a water-based plethysmometer, before and at different times after edema induction. Data were analyzed by ANOVA and Student t test. RESULTS: In the first phase of experiments, some statistically significant effects of homeopathic remedies (Apis, Lachesis and Phosporus) were observed (the reduction in paw volume increase ranging from 10% to 28% at different times since edema induction). In the second phase of experiments, the effects of homeopathic remedies were not confirmed. On the contrary, the unblinded standard allopathic drug indomethacin exhibited its anti-inflammatory effect in both experimental phases (the reduction in paw volume increase ranging from 14% to 40% in the first phase, and from 18% to 38% in the second phase of experiments). CONCLUSION: The discrepancies between single-blind and double-blind methods in animal pharmacological research are noteworthy and should be better investigated, also in non-homeopathic research. [Abstract/Link to Full Text]

Nayak BS, Pinto Pereira LM
Catharanthus roseus flower extract has wound-healing activity in Sprague Dawley rats.
BMC Complement Altern Med. 2006;641.
BACKGROUND: Catharanthus roseus L (C. roseus) has been used to treat a wide assortment of diseases including diabetes. The objective of our study was to evaluate the antimicrobial and wound healing activity of the flower extract of Catharanthus in rats. METHODS: Wound healing activity was determined in rats, after administration (100 mg kg-1 day-1) of the ethanol extract of C. roseus flower, using excision, incision and dead space wounds models. The animals were divided into two groups of 6 each in all the models. In the excision model, group 1 animals were topically treated with carboxymethyl cellulose as placebo control and group 2 received topical application of the ethanol extract of C. roseus at a dose of 100 mg/kg body weight/day. In an incision and dead space model group 1 animals were given normal saline and group 2 received the extract orally at a dose of 100 mg kg-1 day-1. Healing was assessed by the rate of wound contraction, period of epithelization, tensile strength (skin breaking strength), granulation tissue weight, and hydoxyproline content. Antimicrobial activity of the flower extract against four microorganisms was also assessed RESULTS: The extract of C. roseus significantly increased the wound breaking strength in the incision wound model compared with controls (P < 0.001). The extract-treated wounds were found to epithelialize faster, and the rate of wound contraction was significantly increased in comparison to control wounds (P < 0.001), Wet and dry granulation tissue weights, and hydroxyproline content in a dead space wound model increased significantly (p < 0.05). Pseudomonas aeruginosa and Staphylococcus aureus demonstrated sensitivity to C. roseus CONCLUSION: Increased wound contraction and tensile strength, augmented hydroxyproline content along with antimicrobial activity support the use of C. roseus in the topical management of wound healing. [Abstract/Link to Full Text]

Vickers KA, Jolly KB, Greenfield SM
Herbal medicine: women's views, knowledge and interaction with doctors: a qualitative study.
BMC Complement Altern Med. 2006;640.
BACKGROUND: There is growing concern that serious interactions are occurring between prescribed/over the counter and herbal medicines and that there is a lack of disclosure of herbal use by patients to doctors. This study explores women's perspectives about the safety of herbal remedies, herb-drug interactions and communication with doctors about herbal medicines. METHODS: Qualitative, cross-sectional study, with purposive sampling which took place in Cheshire, UK. Eighteen in depth semi-structured interviews were conducted with female herbal medicine users aged 18 years and above. RESULTS: The large majority did not inform their GPs of their use of herbal medicines. This was due to lack of physician enquiry, perception of importance and fear of a negative response. Several women were not aware that herbal remedies could interact with prescribed or over the counter medicines. Of the women who had experienced adverse effects none had reported them, believing them of low importance. CONCLUSION: The women had little knowledge about herb-drug interactions and rarely disclosed use of herbal medicines to their doctor. Doctors' communication and openness regarding herbal medicines needs to improve and there should be increased access to accurate information on herbal medicines in the public and health care domain. [Abstract/Link to Full Text]

Prabuseenivasan S, Jayakumar M, Ignacimuthu S
In vitro antibacterial activity of some plant essential oils.
BMC Complement Altern Med. 2006;639.
BACKGROUND: To evaluate the antibacterial activity of 21 plant essential oils against six bacterial species. METHODS: The selected essential oils were screened against four gram-negative bacteria (Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Proteus vulgaris) and two gram-positive bacteria Bacillus subtilis and Staphylococcus aureus at four different concentrations (1:1, 1:5, 1:10 and 1:20) using disc diffusion method. The MIC of the active essential oils were tested using two fold agar dilution method at concentrations ranging from 0.2 to 25.6 mg/ml. RESULTS: Out of 21 essential oils tested, 19 oils showed antibacterial activity against one or more strains. Cinnamon, clove, geranium, lemon, lime, orange and rosemary oils exhibited significant inhibitory effect. Cinnamon oil showed promising inhibitory activity even at low concentration, whereas aniseed, eucalyptus and camphor oils were least active against the tested bacteria. In general, B. subtilis was the most susceptible. On the other hand, K. pneumoniae exhibited low degree of sensitivity. CONCLUSION: Majority of the oils showed antibacterial activity against the tested strains. However Cinnamon, clove and lime oils were found to be inhibiting both gram-positive and gram-negative bacteria. Cinnamon oil can be a good source of antibacterial agents. [Abstract/Link to Full Text]

Verhoef MJ, Vanderheyden LC, Dryden T, Mallory D, Ware MA
Evaluating complementary and alternative medicine interventions: in search of appropriate patient-centered outcome measures.
BMC Complement Altern Med. 2006;66-38.
BACKGROUND: Central to the development of a sound evidence base for Complementary and Alternative Medicine (CAM) interventions is the need for valid, reliable and relevant outcome measures to assess whether the interventions work. We assessed the specific needs for a database that would cover a wide range of outcomes measures for CAM research and considered a framework for such a database. METHODS: The study was a survey of CAM researchers, practitioners and students. An online questionnaire was emailed to the members of the Canadian Interdisciplinary Network for CAM Research (IN-CAM) and the CAM Education and Research Network of Alberta (CAMera). The majority of survey questions were open-ended and asked about outcome measures currently used, outcome measures' assessment criteria, sources of information, perceived barriers to finding outcome measures and outcome domains of importance. Descriptive quantitative analysis and qualitative content analysis were used. RESULTS: One hundred and sixty-four completed surveys were received. Of these, 62 respondents reported using outcome measures in their CAM research and identified 92 different specific outcomes. The most important barriers were the fact that, for many health concepts, outcome measures do not yet exist, as well as issues related to accessibility of instruments. Important outcome domains identified included physical, psychological, social, spiritual, quality of life and holistic measures. Participants also mentioned the importance of individualized measures that assess unique patient-centered outcomes for each research participant, and measures to assess the context of healing and the process of healing. CONCLUSION: We have developed a preliminary framework that includes all components of health-related outcomes. The framework provides a foundation for a larger, comprehensive collection of CAM outcomes. It fits very well in a whole systems perspective, which requires an expanded set of outcome measures, such as individualized and holistic measures, with attention to issues of process and context. [Abstract/Link to Full Text]

Thompson TD, Weiss M
Homeopathy--what are the active ingredients? An exploratory study using the UK Medical Research Council's framework for the evaluation of complex interventions.
BMC Complement Altern Med. 2006;637.
BACKGROUND: Research in homeopathy has traditionally addressed itself to defining the effectiveness of homeopathic potencies in comparison to placebo medication. There is now increasing awareness that the homeopathic consultation is in itself a therapeutic intervention working independently or synergistically with the prescribed remedy. Our objective was to identify and evalute potential "active ingredients" of the homeopathic approach as a whole, in a prospective formal case series, which draws on actual consultation data, and is based on the MRC framework for the evaluation of complex interventions. METHODS: Following on from a theoretical review of how homeopathic care might mediate its effects, 18 patients were prospectively recruited to a case series based at Bristol Homeopathic Hospital. Patients, who lived with one of three index conditions, were interviewed before and after a five visit "package of care". All consultations were recorded and transcribed verbatim. Additional data, including generic and condition-specific questionnaires, artwork and "significant other" reports were collected. Textual data was subject to thematic analysis and triangulated with other sources. RESULTS: We judged that around one third of patients had experienced a major improvement in their health over the study period, a third had some improvement and a third had no improvement. Putative active ingredients included the patients' "openness to the mind-body connection", consultational empathy, in-depth enquiry into bodily complaints, disclosure, the remedy matching process and, potentially, the homeopathic remedies themselves. CONCLUSION: This study has has identified, using primary consultation and other data, a range of factors that might account for the effectiveness of homeopathic care. Some of these, such as empathy, are non-specific. Others, such as the remedy matching process, are specific to homeopathy. These findings counsel against the use of placebo-controlled RCT designs in which both arms would potentially be receiving specific active ingredients. Future research in homeopathy should focus on pragmatic trials and seek to confirm or refute the therapeutic role of constructs such as patient "openness", disclosure and homeopathicity. [Abstract/Link to Full Text]

Gupta M, Mazumder UK, Gomathi P, Selvan VT
Antiinflammatory evaluation of leaves of Plumeria acuminata.
BMC Complement Altern Med. 2006;636.
BACKROUND: Plumeria acuminata belonging to the family Apocynaceae is commonly known as 'perungalli' in Tamil and is widely distributed throughout the Southern parts of India. In traditional medicinal system different parts of the plant have been mentioned to be useful in a variety of diseases. The plant material is widely used as a purgative, remedy for diarrhoea and cure for itch. The milky juice is employed for the treatment of inflammation and rheumatism. The bark has been reported to be useful in hard tumors, diarrhoea and gonorrhoea. The objective of the present study was to evaluate the antiinflammatory activity of methanol extract of leaves of Plumeria acuminata on carrageenan, dextran, histamine and serotonin-induced inflammation in rat hind paw oedema models. METHODS: Acute and chronic inflammation models were used to evaluate the anti-inflammatory activity of the extract. Wistar albino rats of either sex weighing 180-200 g were used. In acute model carrageenan, dextran, histamine and serotonin models were used to induce inflammation in rat hind paw and cotton pellet-induced granuloma method was used for chronic inflammation model. In each model four groups of six animals were used. In all the models Group I served as control (0.9% normal saline, 5 mlkg(-1) b.w) and group IV as standard (Indomethacin 10 mgkg(-1) b.w). Group II and III received extract at the doses of 250 and 500 mgkg(-1) b.w respectively. RESULTS: The methanol extract of Plumeria acuminata exhibited significant anti-inflammatory activity on the tested experimental animal models. The extract (500 mgkg(-1) b.w) exhibited maximum antiinflammatory effect i.e., 30.51, 47.06, 34.48 and 32.50% (P < 0.001) at the end of 3 h with carrageenan, dextran, histamine and serotonin respectively. Administration of MEPA (500 mgkg(-1) b.w) and indomethacin (10 mgkg(-1) b.w) significantly reduced the formation of granuloma tissue induced by cotton pellet method at a rate of 45.06 and 51.57% respectively. The effect produced by the extract was comparable to that of indomethacin a prototype of a nonsteroidal antiinflammatory agent. CONCLUSION: The results obtained in this study indicated that the methanol extract of Plumeria acuminata possess potent antiinflammatory activity in both acute and chronic models. [Abstract/Link to Full Text]

Duraipandiyan V, Ayyanar M, Ignacimuthu S
Antimicrobial activity of some ethnomedicinal plants used by Paliyar tribe from Tamil Nadu, India.
BMC Complement Altern Med. 2006;635.
BACKGROUND: Antimicrobial activity of 18 ethnomedicinal plant extracts were evaluated against nine bacterial strains (Bacillus subtilis, Staphylococcus aureus, Staphylococcus epidermidis, Enterococcus faecalis, Escherichia coli, Klebsiella pneumonia, Pseudomonas aeruginosa, Ervinia sp, Proteus vulgaris) and one fungal strain (Candida albicans). The collected ethnomedicinal plants were used in folk medicine in the treatment of skin diseases, venereal diseases, respiratory problems and nervous disorders. METHODS: Plants were collected from Palni hills of Southern Western Ghats and the ethnobotanical data were gathered from traditional healers who inhabit the study area. The hexane and methanol extracts were obtained by cold percolation method and the antimicrobial activity was found using paper disc diffusion method. All microorganisms were obtained from Christian Medical College, Vellore, Tamil Nadu, India. RESULTS: The results indicated that out of 18 plants, 10 plants exhibited antimicrobial activity against one or more of the tested microorganisms at three different concentrations of 1.25, 2.5 and 5 mg/disc. Among the plants tested, Acalypha fruticosa, Peltophorum pterocarpum, Toddalia asiatica,Cassia auriculata, Punica granatum and Syzygium lineare were most active. The highest antifungal activity was exhibited by methanol extract of Peltophorum pterocarpum and Punica granatum against Candida albicans. CONCLUSION: This study evaluated the antimicrobial activity of the some ethnomedicinal plants used in folkloric medicine. Compared to hexane extract, methanol extract showed significant activity against tested organisms. This study also showed that Toddalia asiatica, Syzygium lineare, Acalypha fruticosa and Peltophorum pterocarpum could be potential sources of new antimicrobial agents. [Abstract/Link to Full Text]

McEachrane-Gross FP, Liebschutz JM, Berlowitz D
Use of selected complementary and alternative medicine (CAM) treatments in veterans with cancer or chronic pain: a cross-sectional survey.
BMC Complement Altern Med. 2006;634.
BACKGROUND: Complementary and alternative medicine (CAM) is emerging as an important form of care in the United States. We sought to measure the prevalence of selected CAM use among veterans attending oncology and chronic pain clinics and to describe the characteristics of CAM use in this population. METHODS: The self-administered, mail-in survey included questions on demographics, health beliefs, medical problems and 6 common CAM treatments (herbs, dietary supplements, chiropractic care, massage therapy, acupuncture and homeopathy) use. We used the chi-square test to examine bivariate associations between our predictor variables and CAM use. RESULTS: Seventy-two patients (27.3%) reported CAM use within the past 12 months. CAM use was associated with more education (p = 0.02), higher income (p = 0.006), non-VA insurance (p = 0.003), additional care outside the VA (p = 0.01) and the belief that lifestyle contributes to illness (p = 0.015). The diagnosis of chronic pain versus cancer was not associated with differential CAM use (p = 0.15). Seventy-six percent of CAM non-users reported that they would use it if offered at the VA. CONCLUSION: Use of 6 common CAM treatments among these veterans is lower than among the general population, but still substantial. A large majority of veterans reported interest in using CAM modalities if they were offered at the VA. A national assessment of veteran interest in CAM may assist VA leaders to respond to patients' needs. [Abstract/Link to Full Text]

Manna P, Sinha M, Sil PC
Aqueous extract of Terminalia arjuna prevents carbon tetrachloride induced hepatic and renal disorders.
BMC Complement Altern Med. 2006;633.
BACKGROUND: Carbon tetrachloride (CCl4) is a well-known hepatotoxin and exposure to this chemical is known to induce oxidative stress and causes liver injury by the formation of free radicals. Acute and chronic renal damage are also very common pathophysiologic disturbances caused by CCl4. The present study has been conducted to evaluate the protective role of the aqueous extract of the bark of Termnalia arjuna (TA), an important Indian medicinal plant widely used in the preparation of ayurvedic formulations, on CCl4 induced oxidative stress and resultant dysfunction in the livers and kidneys of mice. METHODS: Animals were pretreated with the aqueous extract of TA (50 mg/kg body weight) for one week and then challenged with CCl4 (1 ml/kg body weight) in liquid paraffin (1:1, v/v) for 2 days. Serum marker enzymes, namely, glutamate pyruvate transaminase (GPT) and alkaline phosphatase (ALP) were estimated in the sera of all study groups. Antioxidant status in both the liver and kidney tissues were estimated by determining the activities of the antioxidative enzymes, superoxide dismutase (SOD), catalase (CAT) and glutathione-S-transferase (GST); as well as by determining the levels of thiobarbutaric acid reactive substances (TBARS) and reduced glutathione (GSH). In addition, free radical scavenging activity of the extract was determined from its DPPH radical quenching ability. RESULTS: Results showed that CCl4 caused a marked rise in serum levels of GPT and ALP. TBARS level was also increased significantly whereas GSH, SOD, CAT and GST levels were decreased in the liver and kidney tissue homogenates of CCl4 treated mice. Aqueous extract of TA successfully prevented the alterations of these effects in the experimental animals. Data also showed that the extract possessed strong free radical scavenging activity comparable to that of vitamin C. CONCLUSION: Our study demonstrated that the aqueous extract of the bark of TA could protect the liver and kidney tissues against CCl4-induced oxidative stress probably by increasing antioxidative defense activities. [Abstract/Link to Full Text]

Gauthaman KK, Saleem MT, Thanislas PT, Prabhu VV, Krishnamoorthy KK, Devaraj NS, Somasundaram JS
Cardioprotective effect of the Hibiscus rosa sinensis flowers in an oxidative stress model of myocardial ischemic reperfusion injury in rat.
BMC Complement Altern Med. 2006;632.
BACKGROUND: The present study investigates the cardioprotective effects of Hibiscus rosa sinensis in myocardial ischemic reperfusion injury, particularly in terms of its antioxidant effects. METHODS: The medicinal values of the flowers of Hibiscus rosa sinensis (Chinese rose) have been mentioned in ancient literature as useful in disorders of the heart. Dried pulverized flower of Hibiscus rosa sinensis was administered orally to Wistar albino rats (150-200 gms) in three different doses [125, 250 and 500 mg/kg in 2% carboxy methyl cellulose (CMC)], 6 days per week for 4 weeks. Thereafter, rats were sacrificed; either for the determination of baseline changes in cardiac endogenous antioxidants [superoxide dismutase, reduced glutathione and catalase] or the hearts were subjected to isoproterenol induced myocardial necrosis. RESULTS: There was significant increase in the baseline contents of thiobarbituric acid reactive substances (TBARS) [a measure of lipid per oxidation] with both doses of Hibiscus Rosa sinensis. In the 250 mg/kg treated group, there was significant increase in superoxide dismutase, reduced glutathione, and catalase levels but not in the 125 and 500 mg/kg treated groups. Significant rise in myocardial thiobarbituric acid reactive substances and loss of superoxide dismutase, catalase and reduced glutathione (suggestive of increased oxidative stress) occurred in the vehicle treated hearts subjected to in vivo myocardial ischemic reperfusion injury. CONCLUSION: It may be concluded that flower of Hibiscus rosa sinensis (250 mg/kg) augments endogenous antioxidant compounds of rat heart and also prevents the myocardium from isoproterenol induced myocardial injury. [Abstract/Link to Full Text]

Kwan D, Hirschkorn K, Boon H
U.S. and Canadian pharmacists' attitudes, knowledge, and professional practice behaviors toward dietary supplements: a systematic review.
BMC Complement Altern Med. 2006;631.
BACKGROUND: Although dietary supplements (DS) are widely sold in pharmacies, the legal, ethical, and practice responsibilities of pharmacists with respect to these products have not been well defined. This systematic review of pharmacists' attitudes, knowledge, and professional practice behaviours toward DS is intended to inform pharmacy regulators' and educators' decision making around this topic. METHODS: Eligible studies were identified through a systematic database search for all available years through to March 2006. Articles were analyzed for this review if they included survey data on U.S. or Canadian pharmacists' attitudes, knowledge, or professional practice behaviors toward DS published in 1990 or later. RESULTS: Due to the heterogeneity of the data, it was not possible to draw a conclusion with respect to pharmacists' general attitudes toward DS. Approximately equal numbers of pharmacists report positive as well as negative attitudes about the safety and efficacy of DS. There is strong agreement among pharmacists for the need to have additional training on DS, increased regulation of DS, and quality information on DS. In addition, survey data indicate that pharmacists do not perceive their knowledge of DS to be adequate and that pharmacists do not routinely document, monitor, or inquire about patients' use of DS. Despite this, a large proportion of pharmacists reported receiving questions about DS from patients and other health care practitioners. CONCLUSION: Further research is needed to explore the factors that influence pharmacists' beliefs and attitudes about DS, to accurately evaluate pharmacists' knowledge of DS, and to uncover the reasons why pharmacists do not routinely document, monitor, or inquire about patients' use of DS. [Abstract/Link to Full Text]

Hsieh TC, Wu P, Park S, Wu JM
Induction of cell cycle changes and modulation of apoptogenic/anti-apoptotic and extracellular signaling regulatory protein expression by water extracts of I'm-Yunity (PSP).
BMC Complement Altern Med. 2006;630.
BACKGROUND: I'm-Yunity (PSP) is a mushroom extract derived from deep-layer cultivated mycelia of the patented Cov-1 strain of Coriolus versicolor (CV), which contains as its main bioactive ingredient a family of polysaccharo-peptide with heterogeneous charge properties and molecular sizes. I'm-Yunity (PSP) is used as a dietary supplement by cancer patients and by individuals diagnosed with various chronic diseases. Laboratory studies have shown that I'm-Yunity (PSP) enhances immune functions and also modulates cellular responses to external challenges. Recently, I'm-Yunity (PSP) was also reported to exert potent anti-tumorigenic effects, evident by suppression of cell proliferation and induction of apoptosis in malignant cells. We investigate the mechanisms by which I'm-Yunity (PSP) elicits these effects. METHODS: Human leukemia HL-60 and U-937 cells were incubated with increasing doses of aqueous extracts of I'm-Yunity (PSP). Control and treated cells were harvested at various times and analyzed for changes in: (1) cell proliferation and viability, (2) cell cycle phase transition, (3) induction of apoptosis, (4) expression of cell cycle, apoptogenic/anti-apoptotic, and extracellular regulatory proteins. RESULTS: Aqueous extracts of I'm-Yunity (PSP) inhibited cell proliferation and induced apoptosis in HL-60 and U-937 cells, accompanied by a cell type-dependent disruption of the G1/S and G2/M phases of cell cycle progression. A more pronounced growth suppression was observed in treated HL-60 cells, which was correlated with time- and dose-dependent down regulation of the retinoblastoma protein Rb, diminution in the expression of anti-apoptotic proteins bcl-2 and survivin, increase in apoptogenic proteins bax and cytochrome c, and cleavage of poly(ADP-ribose) polymerase (PARP) from its native 112-kDa form to the 89-kDa truncated product. Moreover, I'm-Yunity (PSP)-treated HL-60 cells also showed a substantial decrease in p65 and to a lesser degree p50 forms of transcription factor NF-kappaB, which was accompanied by a reduction in the expression of cyclooxygenase 2 (COX2). I'm-Yunity (PSP) also elicited an increase in STAT1 (signal transducer and activator of transcription) and correspondingly, decrease in the expression of activated form of ERK (extracellular signal-regulated kinase). CONCLUSION: Aqueous extracts of I'm-Yunity (PSP) induces cell cycle arrest and alterations in the expression of apoptogenic/anti-apoptotic and extracellular signaling regulatory proteins in human leukemia cells, the net result being suppression of proliferation and increase in apoptosis. These findings may contribute to the reported clinical and overall health effects of I'm-Yunity (PSP). [Abstract/Link to Full Text]

Simonsen HT, Adsersen A, Berthelsen L, Christensen SB, Guzmán A, Mřlgaard P
Ethnopharmacological evaluation of radal (leaves of Lomatia hirsuta) and isolation of 2-methoxyjuglone.
BMC Complement Altern Med. 2006;629.
BACKGROUND: Leaves of Lomatia hirsuta are used in traditional medicine in Chile under the common name of "radal". A tea of radal is traditionally used for treatment of cough, bronchial troubles, and asthma. In a preliminary screening, extracts of the leaves revealed antifungal activity, and the present phytochemical study was undertaken to explain this activity and support the traditional use. METHODS: Along with the traditional tea, extracts of the leaves were screened for antifungal and toxic activities. The profile of secondary constituents was obtained using GC-MS. RESULTS: 2-Methoxyjuglone was isolated from the leaves of Lomatia hirsuta and found to be active against the pathogenic fungus Candida albicans (MIC = 8 microg/mL). Cinnamic acid and vanillic acid were identified as major constituents in the tea by GC-MS. The tea was found not to be toxic against Artemia salina. CONCLUSION: The presence of phenolic acids with antimicrobial properties supports the traditional use of Radal, and encourages further studies. [Abstract/Link to Full Text]

Chaya MS, Kurpad AV, Nagendra HR, Nagarathna R
The effect of long term combined yoga practice on the basal metabolic rate of healthy adults.
BMC Complement Altern Med. 2006;628.
BACKGROUND: Different procedures practiced in yoga have stimulatory or inhibitory effects on the basal metabolic rate when studied acutely. In daily life however, these procedures are usually practiced in combination. The purpose of the present study was to investigate the net change in the basal metabolic rate (BMR) of individuals actively engaging in a combination of yoga practices (asana or yogic postures, meditation and pranayama or breathing exercises) for a minimum period of six months, at a residential yoga education and research center at Bangalore. METHODS: The measured BMR of individuals practicing yoga through a combination of practices was compared with that of control subjects who did not practice yoga but led similar lifestyles. RESULTS: The BMR of the yoga practitioners was significantly lower than that of the non-yoga group, and was lower by about 13 % when adjusted for body weight (P < 0.001). This difference persisted when the groups were stratified by gender; however, the difference in BMR adjusted for body weight was greater in women than men (about 8 and 18% respectively). In addition, the mean BMR of the yoga group was significantly lower than their predicted values, while the mean BMR of non-yoga group was comparable with their predicted values derived from 1985 WHO/FAO/UNU predictive equations. CONCLUSION: This study shows that there is a significantly reduced BMR, probably linked to reduced arousal, with the long term practice of yoga using a combination of stimulatory and inhibitory yogic practices. [Abstract/Link to Full Text]

Scheck AC, Perry K, Hank NC, Clark WD
Anticancer activity of extracts derived from the mature roots of Scutellaria baicalensis on human malignant brain tumor cells.
BMC Complement Altern Med. 2006;627.
BACKGROUND: Flavonoid-rich extracts from the mature roots of Scutellaria baicalensis have been shown to exhibit antiproliferative effects on various cancer cell lines. We assessed the ability of an ethanolic extract of S. baicalensis root to inhibit the proliferation of malignant glioma cells. METHODS: Cell lines derived from primary and recurrent brain tumors from the same patient and cells selected for resistance to the chemotherapeutic agent 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU) were used to identify antiproliferative effects of this extract when used alone and in conjunction with BCNU. RESULTS AND DISCUSSION: Results indicated that Scutellaria baicalensis not only inhibits cellular growth in recurrent and drug resistant brain tumor cell lines, but also demonstrates an increased inhibitory effect when used in conjunction with BCNU. CONCLUSION: The results of this study support the efficacy of S. baicalensis as an anticancer agent for glioblastomas multiforme and a potential adjuvant treatment to current chemotherapeutic agents used in the treatment of both primary and recurrent GBMs. Further studies of the effects of individual flavonoids alone and in combination with each other and with currently used therapies are needed. [Abstract/Link to Full Text]

Reisman J, Schachter HM, Dales RE, Tran K, Kourad K, Barnes D, Sampson M, Morrison A, Gaboury I, Blackman J
Treating asthma with omega-3 fatty acids: where is the evidence? A systematic review.
BMC Complement Altern Med. 2006;626.
BACKGROUND: Considerable interest exists in the potential therapeutic value of dietary supplementation with the omega-3 fatty acids. Given the interplay between pro-inflammatory omega-6 fatty acids, and the less pro-inflammatory omega-3 fatty acids, it has been thought that the latter could play a key role in treating or preventing asthma. The purpose was to systematically review the scientific-medical literature in order to identify, appraise, and synthesize the evidence for possible treatment effects of omega-3 fatty acids in asthma. METHODS: Medline, Premedline, Embase, Cochrane Central Register of Controlled Trials, CAB Health, and, Dissertation Abstracts were searched to April 2003. We included randomized controlled trials (RCT's) of subjects of any age that used any foods or extracts containing omega-3 fatty acids as treatment or prevention for asthma. Data included all asthma related outcomes, potential covariates, characteristics of the study, design, population, intervention/exposure, comparators, and co interventions. RESULTS: Ten RCT's were found pertinent to the present report. CONCLUSION: Given the largely inconsistent picture within and across respiratory outcomes, it is impossible to determine whether or not omega-3 fatty acids are an efficacious adjuvant or monotherapy for children or adults. Based on this systematic review we recommend a large randomized controlled study of the effects of high-dose encapsulated omega-3 fatty acids on ventilatory and inflammatory measures of asthma controlling diet and other asthma risk factors. This review was limited because Meta-analysis was considered inappropriate due to missing data; poorly or heterogeneously defined populations, interventions, intervention-comparator combinations, and outcomes. In addition, small sample sizes made it impossible to meaningfully assess the impact on clinical outcomes of co-variables. Last, few significant effects were found. [Abstract/Link to Full Text]

Moffet HH
How might acupuncture work? A systematic review of physiologic rationales from clinical trials.
BMC Complement Altern Med. 2006;625.
BACKGROUND: Scientific interest in acupuncture has led numerous investigators to conduct clinical trials to test the efficacy of acupuncture for various conditions, but the mechanisms underlying acupuncture are poorly understood. METHODS: The author conducted a PubMed search to obtain a fair sample of acupuncture clinical trials published in English in 2005. Each article was reviewed for a physiologic rationale, as well as study objectives and outcomes, experimental and control interventions, country of origin, funding sources and journal type. RESULTS: Seventy-nine acupuncture clinical trials were identified. Twenty-six studies (33%) offered no physiologic rationale. Fifty-three studies (67%) posited a physiologic basis for acupuncture: 33 (62% of 53) proposed neurochemical mechanisms, 2 (4%) segmental nervous system effects, 6 (11%) autonomic nervous system regulation, 3 (6%) local effects, 5 (9%) effects on brain function and 5 (9%) other effects. No rationale was proposed for stroke; otherwise having a rationale was not associated with objective, positive or negative findings, means of intervention, country of origin, funding source or journal type. The dominant explanation for how acupuncture might work involves neurochemical responses and is not reported to be dependent on treatment objective, specific points, means or method of stimulation. CONCLUSION: Many acupuncture trials fail to offer a meaningful rationale, but proposing a rationale can help investigators to develop and test a causal hypothesis, choose an appropriate control and rule out placebo effects. Acupuncture may stimulate self-regulatory processes independent of the treatment objective, points, means or methods used; this would account for acupuncture's reported benefits in so many disparate pathologic conditions. [Abstract/Link to Full Text]

Sherman KJ, Dixon MW, Thompson D, Cherkin DC
Development of a taxonomy to describe massage treatments for musculoskeletal pain.
BMC Complement Altern Med. 2006;624.
BACKGROUND: One of the challenges in conducting research in the field of massage and bodywork is the lack of consistent terminology for describing the treatments given by massage therapists. The objective of this study was to develop a taxonomy to describe what massage therapists actually do when giving a massage to patients with musculoskeletal pain. METHODS: After conducting a review of the massage treatment literature for musculoskeletal pain, a list of candidate techniques was generated for possible inclusion in the taxonomy. This list was modified after discussions with a senior massage therapist educator and seven experienced massage therapists participating in a study of massage for neck pain. RESULTS: The taxonomy was conceptualized as a three level classification system, principal goals of treatment, styles, and techniques. Four categories described the principal goal of treatment (i.e., relaxation massage, clinical massage, movement re-education and energy work). Each principal goal of treatment could be met using a number of different styles, with each style consisting of a number of specific techniques. A total of 36 distinct techniques were identified and described, many of which could be included in multiple styles. CONCLUSION: A new classification system is presented whereby practitioners using different styles of massage can describe the techniques they employ using consistent terminology. This system could help facilitate standardized reporting of massage interventions. [Abstract/Link to Full Text]

Rubio J, Caldas M, Dávila S, Gasco M, Gonzales GF
Effect of three different cultivars of Lepidium meyenii (Maca) on learning and depression in ovariectomized mice.
BMC Complement Altern Med. 2006;623.
BACKGROUND: Lepidium meyenii Walp. (Brassicaceae), known as Maca, is a Peruvian hypocotyl growing exclusively between 4000 and 4500 m altitude in the central Peruvian Andes, particularly in Junin plateau and is used traditionally to enhance fertility. Maca is a cultivated plant and different cultivars are described according to the color of the hypocotyls. METHODS: The study aimed to elucidate the effect of Yellow, Red and Black Maca on cognitive function and depression in ovariectomized (OVX) mice. In all experiments OVX mice were treated during 21 days and divided in four groups: control group, Yellow Maca, Red Maca and Black Maca. Latent learning was assessed using the water finding task and the antidepressant activity of the three varieties of Maca was evaluated using the forced swimming test. Animals were sacrificed at the end of each treatment and the uterus were excised and weighed. RESULTS: Black Maca was the variety that showed the best response in the water finding task, particularly in the trained mice. The three varieties were effective to reduce finding latency in non trained and trained mice (P < 0.05). In the force swimming test, all varieties assessed reduced the time of immobility and increased uterine weight in OVX mice. CONCLUSION: Black Maca appeared to have more beneficial effects on latent learning in OVX mice; meanwhile, all varieties of Maca showed antidepressant activity. [Abstract/Link to Full Text]

Abere TA, Agoreyo FO
Antimicrobial and toxicological evaluation of the leaves of Baissea axillaries Hua used in the management of HIV/AIDS patients.
BMC Complement Altern Med. 2006;622.
BACKGROUND: Persistent diarrhea is a common endemic disease with high incidence among the Africans including Nigerians. It also represents a frequent opportunistic disease in people living with HIV. Diarrhea represents one of the most distressful and persistent symptoms of HIV/AIDS, which may or may not be accompanied by an infection. The leaves decoction of Baissea axillaries Hua (Apocynaceae) is used by traditional herbalists in Edo state, Nigeria for the management of people living with HIV/AIDS. Determination of its antimicrobial activity and toxicological profile will provide supportive scientific evidence in favour of its continuous usage. METHOD: Chemical and chromatographic tests were employed in phytochemical investigations. Inhibitory activities of aqueous and ethanolic extracts against clinical strains of Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus and Streptococcus faecalis were compared with Togamycin (Spectinomycin). Our report includes minimum inhibitory concentration (MIC) against the test organisms. Toxicological evaluation was determined by administering 250 mg/kg and 500 mg/kg of extracts on male Wister rats for 14 days with normal saline as control. The kidneys, liver, heart and testis tissues were examined. RESULTS: Phytochemical studies revealed the presence of alkaloids, tannins, and cyanogenetic glycosides. The extracts inhibited the growth of Escherichia coli, Pseudomonas aeruginosa and Staphylococcus aureus to varying extents, but only the ethanolic extract inhibited growth in Streptococcus faecalis. The LD50 of the extract in mice was above 5000 mg/kg body weight when administered intraperitoneally. Toxicological evaluation showed mere ballooning degeneration of the liver at 250 mg/kg while at 500 mg/kg there was tissue necrosis. The low and high doses showed ill-defined leydig cells in the testis and no remarkable changes in the heart and kidneys. CONCLUSION: Extracts of Baissea axillaries have demonstrated antimicrobial activity against clinical strains of selected microorganisms. While there is toxicity at the dose of 500 mg/kg, the therapy shows potential for application in the treatment of diarrhoea associated with AIDS/HIV. Further studies of Baissea axillaries on diarrhoea and toxicity are necessary to evaluate its mechanism of action and to fully establish its safety profile. [Abstract/Link to Full Text]

Recent Articles in Evidence-based Complementary and Alternative Medicine

Bellavite P, Ortolani R, Pontarollo F, Pitari G, Conforti A
Immunology and Homeopathy. 5. The Rationale of the 'Simile'
Evid Based Complement Alternat Med. 2007 Jun;4(2):149-163.
The foundation of homeopathic medicine is the 'Similia Principle', also known as the 'Principle of Similarity' or also as the 'Simile', which reflects the inversion of pharmacological effects in healthy subjects as compared with sick ones. This article describes the inversion of effects, a widespread medical phenomenon, through three possible mechanisms: non-linearity of dose-response relationship, different initial pathophysiological states of the organism, and pharmacodynamics of body response to the medicine. Based on the systemic networks which play an important role in response to stress, a unitary and general model is designed: homeopathic medicines could interact with sensitive (primed) regulation systems through complex information, which simulate the disorders of natural disease. Reorganization of regulation systems, through a coherent response to the medicine, could pave the way to the healing of the cellular, tissue and neuro-immuno-endocrine homeodynamics. Preliminary evidence is suggesting that even ultra-low doses and high-dilutions of drugs may incorporate structural or frequency information and interact with chaotic dynamics and physical-electromagnetic levels of regulation. From the clinical standpoint, the 'simile' can be regarded as a heuristic principle, according to which the detailed knowledge of pathogenic effects of drugs, associated with careful analysis of signs and symptoms of the ill subject, could assist in identifying homeopathic remedies with high grade of specificity for the individual case. [Abstract/Link to Full Text]

Joshi K, Hankey A, Patwardhan B
Traditional phytochemistry: identification of drug by 'taste'.
Evid Based Complement Alternat Med. 2007 Jun;4(2):145-8.
Ayurveda, the system of traditional medicine from India, holds that 'Rasa', a concept roughly corresponding to taste, is a basis for identifying pharmacological properties of plants and other materia medica used in Dravyaguna-its system of phytomedicine. This idea has recently found support in studies of ibuprofen, the pharmacological properties of which are similar to those of oleocanthal, because the two substances have very similar tastes. This paper discusses a possible scientific approach to understanding the Ayurvedic (hypo)thesis in terms of the stereochemical basis of both pharamaco-activity and taste, and the numbers of possible pharmaco-active compounds that 'Rasa' may be able to distinguish. We conclude that molecules binding to a specific enzyme active site should have their own 'Rasa', and that the number of different subjectively experienced 'tastes' is more than enough to distinguish between molecular shapes binding to all enzyme active sites in the body. [Abstract/Link to Full Text]

Hankey A, Ewing E
New Light on Chromotherapy: Grakov's 'Virtual Scanning' System of Medical Assessment and Treatment.
Evid Based Complement Alternat Med. 2007 Jun;4(2):139-44.
Virtual Scanning incorporates novel uses of colored light into its system of health assessment and therapy. Independent investigations of its effectiveness in Russia and the UK have revealed unique abilities to correct incipient and fully developed chronic conditions. As such it forms an important new addition to the field of Chromotherapy. It differs from most others, in that its development depended on discoveries in neuroscience by its inventor, and subsequent application of new models in computational neuroscience. [Abstract/Link to Full Text]

Cooper EL
Is There Room for Paradox in CAM?
Evid Based Complement Alternat Med. 2007 Jun;4(2):135-7. [Abstract/Link to Full Text]

Hankey A
CAM and Post-Traumatic Stress Disorder.
Evid Based Complement Alternat Med. 2007 Mar;4(1):131-2.
In the form of the Transcendental Meditation program CAM offers a method of eliminating deep-rooted stress, the efficacy of which has been demonstrated in several related studies. Any discussion of CAM and post-traumatic stress disorder should include a study of its application to Vietnam War Veterans in which improvements were observed on all variables, and several participants were able to return to work after several years of being unable to hold a job. The intervention has been studied for its impact on brain and autonomic nervous system function. It has been found to be highly effective against other stress-related conditions such as hypertension, and to improve brain coherence-a measure of effective brain function. It should be considered a possible 'new and improved mode of treatment' for PTSD, and further studies of its application made. [Abstract/Link to Full Text]

Qu F, Zhou J
Electro-acupuncture in relieving labor pain.
Evid Based Complement Alternat Med. 2007 Mar;4(1):125-30.
To study the efficacy of electro-acupuncture for the relief of labor pain, and to build a better understanding of how electro-acupuncture might influence the neuroendocrine system, 36 primiparas were randomly divided into an electro-acupuncture group and a control group. Assessments of pain intensity and degree of relaxation during labor were analyzed. The differences between the electro-acupuncture group and the control group on the concentration of beta-endorphin (beta-EP) and 5-hydroxytryptamine (5-HT) in the peripheral blood were compared. The electro-acupuncture group was found to exhibit a lower pain intensity and a better degree of relaxation than the control group (p = 0.018; p = 0.031). There existed a significant difference in the concentration of beta-EP and 5-HT in the peripheral blood between the two groups at the end of the first stage (p = 0.037; p = 0.030). Electro-acupuncture was found to be an effective alternative or complementary therapy in the relief of pain during labor. The benefit of electro-acupuncture for relieving labor pain may be based on the mechanism of producing a synergism of the central nervous system (CNS) with a direct impact on the uterus through increasing the release of beta-EP and 5-HT into the peripheral blood. [Abstract/Link to Full Text]

Weze C, Leathard HL, Grange J, Tiplady P, Stevens G
Healing by Gentle Touch Ameliorates Stress and Other Symptoms in People Suffering with Mental Health Disorders or Psychological Stress.
Evid Based Complement Alternat Med. 2007 Mar;4(1):115-123.
Previous studies on healing by gentle touch in clients with various illnesses indicated substantial improvements in psychological well-being, suggesting that this form of treatment might be helpful for people with impaired quality of mental health. The purpose of this study was to evaluate the effectiveness and safety of healing by gentle touch in subjects with self-reported impairments in their psychological well-being or mental health. One hundred and forty-seven clients who identified themselves as having psychological problems received four treatment sessions. Pre- to post-treatment changes in psychological and physical functioning were assessed by self-completed questionnaires which included visual analogue scales (VAS) and the EuroQoL (EQ-5D). Participants recorded reductions in stress, anxiety and depression scores and increases in relaxation and ability to cope scores (all P < 0.0004). Improvements were greatest in those with the most severe symptoms initially. This open study provides strong circumstantial evidence that healing by gentle touch is safe and effective in improving psychological well-being in participants with self-reported psychological problems, and also that it safely complements standard medical treatment. Controlled trials are warranted. [Abstract/Link to Full Text]

Abbott RB, Hui KK, Hays RD, Li MD, Pan T
A Randomized Controlled Trial of Tai Chi for Tension Headaches.
Evid Based Complement Alternat Med. 2007 Mar;4(1):107-113.
This study examined whether a traditional low-impact mind-body exercise, Tai Chi, affects health-related quality-of-life (HRQOL) and headache impact in an adult population suffering from tension-type headaches. Forty-seven participants were randomly assigned to either a 15 week intervention program of Tai Chi instruction or a wait-list control group. HRQOL (SF-36v2) and headache status (HIT-6trade mark) were obtained at baseline and at 5, 10 and 15 weeks post-baseline during the intervention period. Statistically significant (P < 0.05) improvements in favor of the intervention were present for the HIT score and the SF-36 pain, energy/fatigue, social functioning, emotional well-being and mental health summary scores. A 15 week intervention of Tai Chi practice was effective in reducing headache impact and also effective in improving perceptions of some aspects of physical and mental health. [Abstract/Link to Full Text]

Taniguchi N, Kanai S
Efficacy of Static Magnetic Field for Locomotor Activity of Experimental Osteopenia.
Evid Based Complement Alternat Med. 2007 Mar;4(1):99-105.
In order to examine the effectiveness of applying a static magnetic field (SMF) for increasing bone mineral density (BMD), we assessed the degree of osteopenia by dual-energy X-ray absorptiometry (DEXA), the metabolism measuring system, and histological examination of bone tissue in an ovariectomized (OVX) rat model. Thirty-six female Wistar rats (8 weeks old, 160-180 g) were divided into three groups. The rats in the OVX-M group were exposed to SMF for 12 weeks after ovariectomy. The ovariectomized rats in the OVX-D group were not exposed to SMF as a control. The rats in the normal group received neither ovariectomy nor exposure to SMF. Twelve-week exposure to SMF in the OVX-M group inhibited the reduction in BMD that was observed in the OVX-D group. Moreover, in the OVX rats, before exposure to SMF, there was no clear difference in the level of locomotor activity between the active and resting phases, and the pattern of locomotor activity was irregular. After exposure of OVX rats to SMF, the pattern of locomotor activity became diphasic with clear active and resting phases, as was observed in the normal group. In the OVX-M group, the continuity of the trabecular bone was maintained more favorably and bone mass was higher than the respective parameters in the OVX-D group. These results demonstrate that exposure to SMF increased the level of locomotor activity in OVX rats, thereby increasing BMD. [Abstract/Link to Full Text]

Moghaddam FM, Farimani MM, Salahvarzi S, Amin G
Chemical Constituents of Dichloromethane Extract of Cultivated Satureja khuzistanica.
Evid Based Complement Alternat Med. 2007 Mar;4(1):95-8.
Four compounds beta-sitosterol, beta-sitosterol-3-O-beta-d-glucopyranoside, ursolic acid and 4',5,6-trihydroxy-3', 7-dimethoxyflavone were characterized from the dichloromethane extract of the aerial parts of Satureja khuzistanica (Lamiaceae), a native medicinal plant growing in Iran, on the basis of spectral analysis and comparing with the data in literature. The natural occurrence of these compounds can be conclusive for the chemotaxonomic characterization of the genus Satureja. [Abstract/Link to Full Text]

Kawamura M, Kasai H
Delayed Cell Cycle Progression and Apoptosis Induced by Hemicellulase-Treated Agaricus blazei.
Evid Based Complement Alternat Med. 2007 Mar;4(1):83-94.
We examined the effects of hemicellulase-treated Agaricus blazei (AB fraction H, ABH) on growth of several tumor cell lines. ABH inhibited the proliferation of some cell lines without cytotoxic effects. It markedly prolonged the S phase of the cell cycle. ABH also induced mitochondria-mediated apoptosis in different cell lines. However, it had no impact on the growth of other cell lines. ABH induced strong activation of p38 mitogen-activated protein kinase (MAPK) in the cells in which it evoked apoptosis. On the other hand, ABH showed only a weak p38 activation effect in those cell lines in which it delayed cell cycle progression with little induction of apoptosis. However, p38 MAPK-specific inhibitor inhibited both ABH-induced effects, and ABH also caused apoptosis in the latter cells under conditions of high p38 MAPK activity induced by combined treatment with TNF-alpha. These results indicate that the responsiveness of p38 MAPK to ABH, which differs between cell lines, determines subsequent cellular responses on cell growth. [Abstract/Link to Full Text]

Johng HM, Yoo JS, Yoon TJ, Shin HS, Lee BC, Lee C, Lee JK, Soh KS
Use of magnetic nanoparticles to visualize threadlike structures inside lymphatic vessels of rats.
Evid Based Complement Alternat Med. 2007 Mar;4(1):77-82.
A novel application of fluorescent magnetic nanoparticles was made to visualize a new tissue which had not been detectable by using simple stereomicroscopes. This unfamiliar threadlike structure inside the lymphatic vessels of rats was demonstrated in vivo by injecting nanoparticles into lymph nodes and applying magnetic fields on the collecting lymph vessels so that the nanoparticles were taken up by the threadlike structures. Confocal laser scanning microscope images of cryosectioned specimens exhibited that the nanoparticles were absorbed more strongly by the threadlike structure than by the lymphatic vessels. Further examination using a transmission electron microscope revealed that the nanoparticles had been captured between the reticular fibers in the extracellular matrix of the threadlike structures. The emerging technology of nanoparticles not only allows the extremely elusive threadlike structures to be visualized but also is expected to provide a magnetically controllable means to investigate their physiological functions. [Abstract/Link to Full Text]

Liu HX, Tian JB, Luo F, Jiang YH, Deng ZG, Xiong L, Liu C, Wang JS, Han JS
Repeated 100 Hz TENS for the Treatment of Chronic Inflammatory Hyperalgesia and Suppression of Spinal Release of Substance P in Monoarthritic Rats.
Evid Based Complement Alternat Med. 2007 Mar;4(1):65-75.
Transcutaneous electrical nerve stimulation (TENS) has been shown to be an effective measure for pain relief. The aim of the present study was to determine the optimal intensity and interval of repeated 100 Hz TENS for the treatment of chronic inflammatory hyperalgesia in a monoarthritic pain model of the rat, and to assess the changes of the spinal substance P (SP) release in response to TENS treatment. A reliable, reproducible chronic monoarthritic pain model was produced by intra-articular injection of complete Freund's adjuvant (CFA) at single ankle joint. The efficacy of 100 Hz TENS treatments with different frequencies and intensities was compared. In the acute period (within 3 weeks) of monoarthritis, twice-a-week schedule of TENS reduced the swelling of the inflamed ankle significantly. In the stable period (4-9 weeks), however, once-a-week schedule produced a significantly better therapeutic effect on both inflammation and arthritic hyperalgesia than that of twice- or five-times-a-week schedule. Using three levels of intensity of TENS, we found that the weaker (1-1-2 mA) stimulation produced significantly better therapeutic effects. Repeated TENS produced a reduction of SP content in spinal perfusate in parallel with the progressive reduction of the arthritic pain scores. Our results suggest that (i) consecutive TENS treatments produced cumulative effect for chronic hyperalgesia, (ii) for chronic inflammatory hyperalgesia, a weaker intensity and more sparsely arranged treatment schedule may produce better therapeutic effect and (iii) a decrease in SP release may serve as one of the possible neurochemical mechanisms underlying the therapeutic effects of multiple TENS treatments on chronic inflammatory hyperalgesia. [Abstract/Link to Full Text]

Xu S, Tomita N, Ikeuchi K, Ikada Y
Recovery of Small-Sized Blood Vessels in Ischemic Bone Under Static Magnetic Field.
Evid Based Complement Alternat Med. 2007 Mar;4(1):59-63.
Effects of static magnetic field (SMF) on the vascularization in bone were evaluated using an ischemic bone model, where rat femoral artery was ligated. Magnetized and unmagnetized samarium-cobalt rods were implanted transcortically into the middle diaphysis of the ischemic femurs. Collateral circulation was evaluated by injection of microspheres into the abdominal aorta at the third week after ligation. It was found that the bone implanted with a magnetized rod showed a larger amount of trapped microspheres than that with an unmagnetized rod at the proximal and the distal region (P < 0.05 proximal region). There were no significant differences at the middle and the distal region. This tendency was similar to that of the bone mineral density in the SMF-exposed ischemic bone. [Abstract/Link to Full Text]

Yim YK, Lee H, Hong KE, Kim YI, Lee BR, Son CG, Kim JE
Electro-acupuncture at acupoint ST36 reduces inflammation and regulates immune activity in Collagen-Induced Arthritic Mice.
Evid Based Complement Alternat Med. 2007 Mar;4(1):51-7.
This study aimed to investigate the anti-inflammatory, anti-arthritic and immuno-regulatory effects of electro-acupuncture (EA) at ST36 on Collagen-induced arthritis (CIA) in mice. Male DBA/1J mice were divided into five groups: Normal, Control, NR (needle retention), EAI and EAII. All mice except those in the normal group were immunized with Collagen II for arthritis induction. Acupuncture needles were inserted into mice ST36 and electrical currents at a frequency of 2 Hz in a continuous rectangular wave form were conducted through the needles for 15 min, 3 times a week. EA treatments were administered for 5 weeks in the EAI group and for 9 weeks in the EAII group. The mice in the NR group were acupunctured in the same manner as the EA groups and the needles were retained for 15 min without electrical stimulation. CIA incidence analysis, ELISA, histological analysis and FACS analysis were performed to evaluate the effect of EA on CIA. EA at ST36 significantly reduced CIA incidence, IL-6, TNF-a, INF-gamma, collagen II antibody, IgG and IgM levels in CIA mice serum and prevented knee joint destruction. EA at ST36 also reduced CD69+/CD3e+ cells and CD11a+/CD19+ cells in CIA mice lymph nodes, and CD11b+/Gr1+ cells in CIA mice knee joints. The ratios of CD3e+ cells to CD19+ cells, and CD8+ cells to CD4+ cells were maintained closer to the normal range in the EA groups as compared with the control group or the NR group. EAII was more effective than EAI throughout all the measurements. The NR was effective as well, though less effective than EA. EA at ST36 may have an anti-inflammatory, anti-arthritic and immuno-regulatory effects on CIA in mice. The effectiveness is stronger when EA starts earlier and is applied longer. Needle retention without electrical stimulation may be effective on CIA as well, however less effective than EA. Electrical stimulation and acupoint ST36 may have synergistic effects on CIA. [Abstract/Link to Full Text]

Nakhai LA, Mohammadirad A, Yasa N, Minaie B, Nikfar S, Ghazanfari G, Zamani MJ, Dehghan G, Jamshidi H, Boushehri VS, Khorasani R, Abdollahi M
Benefits of Zataria multiflora Boiss in Experimental Model of Mouse Inflammatory Bowel Disease.
Evid Based Complement Alternat Med. 2007 Mar;4(1):43-50.
Inflammatory bowel disease (IBD) is a chronic condition of the intestine with unknown etiology involving multiple immune, genetic and environmental factors. We were interested to examine the effect of total extract from Zataria multiflora Boiss, a folk medicinal plant on prevention and treatment of experimental IBD. Z. multiflora was administered (400, 600, 900 p.p.m.) through drinking water to IBD mice induced by intrarectal administration of acetic acid. Prednisolone was used as the standard drug for comparison. Biochemical, macroscopic and microscopic examinations of colon were performed. Biochemical evaluation of inflamed colon was done using assay of myeloperoxidase (MPO) activity and thiobarbituric acid reactive substances (TBARS) concentration as indicators of free radical activity and cell lipid peroxidation. The activity of MPO and lipid peroxidation products (TBARS) increased in acetic acid-treated groups while recovered by pretreatment of animals with Z. multiflora (400-900 p.p.m.) and prednisolone. Z. multiflora (600 and 900 p.p.m.) and prednisolone-treated groups showed significantly lower score values of macroscopic and microscopic characters when compared with the acetic acid-treated group. The beneficial effect of Z. multiflora (900 p.p.m.) was comparable with that of prednisolone. The antioxidant, antimicrobial and anti-inflammatory potentials of Z. multiflora might be the mechanisms by which this herbal extract protects animals against experimentally induced IBD. Proper clinical investigation should be carried out to confirm the activity in human. [Abstract/Link to Full Text]

Bhat MS, Rao G, Murthy KD, Bhat PG
Housing in pyramid counteracts neuroendocrine and oxidative stress caused by chronic restraint in rats.
Evid Based Complement Alternat Med. 2007 Mar;4(1):35-42.
The space within the great pyramid and its smaller replicas is believed to have an antistress effect. Research has shown that the energy field within the pyramid can protect the hippocampal neurons of mice from stress-induced atrophy and also reduce neuroendocrine stress, oxidative stress and increase antioxidant defence in rats. In this study, we have, for the first time, attempted to study the antistress effects of pyramid exposure on the status of cortisol level, oxidative damage and antioxidant status in rats during chronic restraint stress. Adult female Wistar rats were divided into four groups as follows: normal controls (NC) housed in home cage and left in the laboratory; restrained rats (with three subgroups) subject to chronic restraint stress by placing in a wire mesh restrainer for 6 h per day for 14 days, the restrained controls (RC) having their restrainers kept in the laboratory; restrained pyramid rats (RP) being kept in the pyramid; and restrained square box rats (RS) in the square box during the period of restraint stress everyday. Erythrocyte malondialdehyde (MDA) and plasma cortisol levels were significantly increased and erythrocyte-reduced glutathione (GSH) levels, erythrocyte glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) activities were significantly decreased in RC and RS rats as compared to NC. However, these parameters were maintained to near normal levels in RP rats which showed significantly decreased erythrocyte MDA and plasma cortisol and significantly increased erythrocyte GSH levels, erythrocyte GSH-Px and SOD activities when compared with RS rats. The results showed that housing in pyramid counteracts neuroendocrine and oxidative stress caused by chronic restraint in rats. [Abstract/Link to Full Text]

Beider S, Moyer CA
Randomized controlled trials of pediatric massage: a review.
Evid Based Complement Alternat Med. 2007 Mar;4(1):23-34.
The existing reviews of massage therapy (MT) research are either limited to infants, adults, or were conducted prior to the publication of the most recent studies using pediatric samples. Randomized controlled trials (RCTs) of pediatric MT are reviewed. A literature search yielded 24 RCTs of pediatric MT, defined as the manual manipulation of soft tissue intended to promote health and well-being in recipients between 2 and 19 years of age. Because RCTs of pediatric MT varied considerably in the amount and types of data reported, quantitative and narrative review methods were both used. Single-dose and multiple-dose effects were examined separately. Among single-dose effects, significant reductions of state anxiety were observed at the first session (g = 0.59, P < 0.05) and the last session (g = 1.10, P < 0.01) of a course of treatment. Effects for salivary cortisol (g = 0.28), negative mood (g = 0.52) and behavior (g = 0.37) were non-significant. Three of eleven multiple-dose effects were statistically significant. These were trait anxiety (g = 0.94, P < 0.05), muscle tone (g = 0.90, P < 0.01) and arthritis pain (g = 1.33, P < 0.01). Results of studies not permitting effect size calculation were judged to be generally consistent with quantitative results. MT benefits pediatric recipients, though not as universally as sometimes reported. Numerous weaknesses endemic to MT research (e.g. low statistical power, frequent failure to report basic descriptive statistics) are identified, and recommendations for future pediatric MT research are discussed. [Abstract/Link to Full Text]

Liang W, Binns CW, Jian L, Lee AH
Does the consumption of green tea reduce the risk of lung cancer among smokers?
Evid Based Complement Alternat Med. 2007 Mar;4(1):17-22.
Experimental and epidemiological studies were reviewed to assess whether the consumption of green tea could reduce the risk of lung cancer in smokers. Articles published since 1990 were located by searching electronic databases PubMed, Ovid and Science Direct, using keywords 'lung cancer', 'tea' and 'smoking' without any restriction on language. After relevant articles had been located, further papers were obtained from their reference lists. Evidence from experimental studies (in vitro animal and human trials) suggested that regular intake of green tea may be protective against tobacco carcinogens. However, the mechanism behind the protective effect is only partly understood. In most of the epidemiological studies reviewed, the green tea exposure was within 5 years of the interview or follow-up, which would coincide with the induction period and latent period of lung cancer. Longer term studies are thus needed to further quantify the cancer risk. There is some evidence suggesting regular intake of green tea at high level (>3 cups per day) may reduce the risk of smokers developing lung cancer. Improvement in measuring green tea intake is required in order to confirm the evidence from epidemiological studies. [Abstract/Link to Full Text]

Milgrom LR
Journeys in the country of the blind: entanglement theory and the effects of blinding on trials of homeopathy and homeopathic provings.
Evid Based Complement Alternat Med. 2007 Mar;4(1):7-16.
The idea of quantum entanglement is borrowed from physics and developed into an algebraic argument to explain how double-blinding randomized controlled trials could lead to failure to provide unequivocal evidence for the efficacy of homeopathy, and inability to distinguish proving and placebo groups in homeopathic pathogenic trials. By analogy with the famous double-slit experiment of quantum physics, and more modern notions of quantum information processing, these failings are understood as blinding causing information loss resulting from a kind of quantum superposition between the remedy and placebo. [Abstract/Link to Full Text]

Liu T
Role of acupuncturists in acupuncture treatment.
Evid Based Complement Alternat Med. 2007 Mar;4(1):3-6.
Traditional Chinese medicine (TCM) is based on a paradigm of the body different to that of modern biomedicine. Inherent characteristics of TCM necessitate an active and central role of acupuncturists in acupuncture treatment. The author looks at acupuncture in the practical context and analyzes the role of acupuncturists in diagnostic process and treatment delivery. Acupuncture as a complex non-pharmacological therapy depends solely on the acupuncturists' skills, competence and understanding of TCM theory to work. More attention should be given to this important role of acupuncturists in either clinical practice or research on acupuncture. [Abstract/Link to Full Text]

Cooper EL
On the Road to an Impact Factor for eCAM.
Evid Based Complement Alternat Med. 2007 Mar;4(1):1-2. [Abstract/Link to Full Text]

Evidence-based Complementary and Alternative Medicine.
Evid Based Complement Alternat Med. 2006 Dec;3(4):555-8. [Abstract/Link to Full Text]

Flowers J
What is qi?
Evid Based Complement Alternat Med. 2006 Dec;3(4):551-2. [Abstract/Link to Full Text]

Marotta F
Complementary and alternative medicine in clinical practice. Complementary and allopathic medicine: the hospital as a place of possible integration: december 2-3, 2005, conference room, s. Carlo borromeo hospital, milan.
Evid Based Complement Alternat Med. 2006 Dec;3(4):549-50. [Abstract/Link to Full Text]

Epstein OI, Pavlov IF, Shtark MB
Improvement of Memory by Means of Ultra-Low Doses of Antibodies to S-100B Antigen.
Evid Based Complement Alternat Med. 2006 Dec;3(4):541-5.
Antigen S-100B of nervous tissue, according to the data of numerous studies, affects the mechanisms of nervous system plasticity and memory. The influence of ultralow doses of antibodies to S-100B (6C dilution, according to the homeopathic pharmacopoeia) has been studied on three learning behavioral models on Wistar rats, which were inhibitory avoidance, choosing of bowls with sucrose and feeding behavior cessation after auditory signal. For all three tasks, parameters of reproduction of the learned skills improved after per oral administration of potentiated antibodies to S-100B antigen immediately after learning. Possible mechanisms of the anti-S-100B antibodies influence on memory formation are discussed. [Abstract/Link to Full Text]

Canter PH, Brown LB, Greaves C, Ernst E
Johrei family healing: a pilot study.
Evid Based Complement Alternat Med. 2006 Dec;3(4):533-40.
Johrei is a form of spiritual healing comprising "energy channelling" and light massage given either by a trained healer or, after some basic training, by anyone. This pilot trial aimed to identify any potential benefits of family-based Johrei practice in childhood eczema and for general health and to establish the feasibility of a subsequent randomised controlled trial. Volunteer families of 3-5 individuals, including at least one child with eczema were recruited to an uncontrolled pilot trial lasting 12 months. Parents were trained in Johrei healing and then practised at home with their family. Participants kept diaries and provided questionnaire data at baseline, 3,6 and 12 months. Eczema symptoms were scored at the same intervals. Scepticism about Johrei is presently an obstacle to recruitment and retention of a representative sample in a clinical trial, and to its potential use in general practice. The frequency and quality of practise at home by families may be insufficient to bring about the putative health benefits. Initial improvements in eczema symptoms and diary recorded illness, could not be separated from seasonal factors and other potential confounders. There were no improvements on other outcomes measuring general health and psychological wellbeing of family members. [Abstract/Link to Full Text]

Carlsson M, Arman M, Backman M, Flatters U, Hatschek T, Hamrin E
A Five-year Follow-up of Quality of Life in Women with Breast Cancer in Anthroposophic and Conventional Care.
Evid Based Complement Alternat Med. 2006 Dec;3(4):523-31.
Complementary and alternative medicine is used by many cancer patients in most parts of the world, and its use is increasing. The aim of the present study was to examine, over 5 years, the perceived quality of life/life satisfaction in two samples of women with breast cancer who were treated with anthroposophic care or conventional medical treatment only. Data from admission, after 1 year and after 5 years are used for the comparisons. On admission to the study the women in anthroposophic care perceived their quality of life to be lower than that of the women in the conventional treatment group, especially for emotional, cognitive and social functioning and overall quality of life. Sixty women who actively chose treatment with anthroposophic medicine and 60 individually matched women treated with conventional medicine participated. Quality of life was measured by the EORTC QLQ-C30 and the Life Satisfaction Questionnaire. Twenty-six women within anthroposophic care and 31 women within conventional medicine survived the 5 years. Effect size (ES) estimation favored the anthroposophic group in seven of the subscales mostly measuring emotional functioning. The ES for four of the subscales favored the conventional treatment group, mostly concerning physical functioning. After 5 years there were improvements in overall quality of life and in emotional and social functioning compared to admission for the women in anthroposophic care. The improvements took place between admission and 1 year, but not further on. Only minor improvements were found in the matching group. [Abstract/Link to Full Text]

Hankey A
Studies of advanced stages of meditation in the tibetan buddhist and vedic traditions. I: a comparison of general changes.
Evid Based Complement Alternat Med. 2006 Dec;3(4):513-21.
This article is the first of two comparing findings of studies of advanced practitioners of Tibetan Buddhist meditation in remote regions of the Himalayas, with established results on long-term practitioners of the Transcendental Meditation programs. Many parallel levels of improvement were found, in sensory acuity, perceptual style and cognitive function, indicating stabilization of aspects of attentional awareness. Together with observed increases in EEG coherence and aspects of brain function, such changes are consistent with growth towards a state of total brain functioning, i.e. development of full mental potential. They are usually accompanied by improved health parameters. How they may be seen to be consistent with growth of enlightenment will be the subject of a second article. [Abstract/Link to Full Text]

Chawla R, Arora R, Singh S, Sagar RK, Sharma RK, Kumar R, Sharma A, Tripathi RP, Puri SC, Khan HA, Shawl AS, Sultan P, Krishan T, Qazi GN
Podophyllum hexandrum Offers Radioprotection by Modulating Free Radical Flux: Role of Aryl-Tetralin Lignans.
Evid Based Complement Alternat Med. 2006 Dec;3(4):503-11.
We have evaluated the effect of variation in aryl-tetralin lignans on the radioprotective properties of Podophyllum hexandrum. Two fractionated fractions of P. hexandrum [methanolic (S1) and chloroform fractions (S2)], with varying aryl-tetralin lignan content were utilized for the present study. The peroxyl ion scavenging potentials of S1 and S2 were found to be comparable [i.e. 45.88% (S1) and 41% (S2)] after a 48 h interval in a time-dependent study, whereas in a 2 h study, S2 exhibited significant (P < 0.05) antioxidant activity in different metal ion + flux states. In the aqueous phase, S2 exhibited non-site-specific reactive oxygen species scavenging activity, i.e. 73.12% inhibition at 500 mug ml(-1). S1 exhibited 58.40 +/- 0.8% inhibition (at 0.025 mug ml(-1)) of the formation of reactive nitrite radicals, comparable to S2 (52.45 +/- 0.825%), and also showed 45.01% site-specific activity (1000 mug ml(-1)), along with significant (P < 0.05) electron donation potential (50-2000 mug ml(-1)) compared to S2. Such activities of S1 could be attributed to the significantly (P < 0.05) higher levels of podophyllotoxin beta-d-glucopyranoside (16.5 times) and demethyl podophyllotoxin glucoside (2.9 times) compared with S2. Together, these findings clearly prove that aryl-tetralin lignan content influences the radiation protective potential of the Podophyllum fractions to a great extent. [Abstract/Link to Full Text]

Recent Articles in Journal of Negative Results in Biomedicine

Hollox EJ, Davies J, Griesenbach U, Burgess J, Alton EW, Armour JA
Beta-defensin genomic copy number is not a modifier locus for cystic fibrosis.
J Negat Results Biomed. 2005;49.
Human beta-defensin 2 (DEFB4, also known as DEFB2 or hBD-2) is a salt-sensitive antimicrobial protein that is expressed in lung epithelia. Previous work has shown that it is encoded in a cluster of beta-defensin genes at 8p23.1, which varies in copy number between 2 and 12 in different individuals. We determined the copy number of this locus in 355 patients with cystic fibrosis (CF), and tested for correlation between beta-defensin cluster genomic copy number and lung disease associated with CF. No significant association was found. [Abstract/Link to Full Text]

Wagenleiter SE, Jagiello P, Akkad DA, Arning L, Griga T, Klein W, Epplen JT
On the genetic involvement of apoptosis-related genes in Crohn's disease as revealed by an extended association screen using 245 markers: no evidence for new predisposing factors.
J Negat Results Biomed. 2005;48.
Crohn's disease (CD) presents as an inflammatory barrier disease with characteristic destructive processes in the intestinal wall. Although the pathomechanisms of CD are still not exactly understood, there is evidence that, in addition to e.g. bacterial colonisation, genetic predisposition contributes to the development of CD. In order to search for predisposing genetic factors we scrutinised 245 microsatellite markers in a population-based linkage mapping study. These microsatellites cover gene loci the encoded protein of which take part in the regulation of apoptosis and (innate) immune processes. Respective loci contribute to the activation/suppression of apoptosis, are involved in signal transduction and cell cycle regulators or they belong to the tumor necrosis factor superfamily, caspase related genes or the BCL2 family. Furthermore, several cytokines as well as chemokines were included. The approach is based on three steps: analyzing pooled DNAs of patients and controls, verification of significantly differing microsatellite markers by genotyping individual DNA samples and, finally, additional reinvestigation of the respective gene in the region covered by the associated microsatellite by analysing single-nucleotide polymorphisms (SNPs). Using this step-wise process we were unable to demonstrate evidence for genetic predisposition of the chosen apoptosis- and immunity-related genes with respect to susceptibility for CD. [Abstract/Link to Full Text]

Gödde R, Brune S, Jagiello P, Sindern E, Haupts M, Schimrigk S, Müller N, Epplen JT
An extended association screen in multiple sclerosis using 202 microsatellite markers targeting apoptosis-related genes does not reveal new predisposing factors.
J Negat Results Biomed. 2005;47.
Apoptosis, the programmed death of cells, plays a distinct role in the etiopathogenesis of Multiple sclerosis (MS), a common disease of the central nervous system with complex genetic background. Yet, it is not clear whether the impact of apoptosis is due to altered apoptotic behaviour caused by variations of apoptosis-related genes. Instead, apoptosis in MS may also represent a secondary response to cellular stress during acute inflammation in the central nervous system. Here, we screened 202 apoptosis-related genes for association by genotyping 202 microsatellite markers in initially 160 MS patients and 160 controls, both divided in 4 sets of pooled DNA samples, respectively. When applying Bonferroni correction, no significant differences in allele frequencies were detected between MS patients and controls. Nevertheless, we chose 7 markers for retyping in individual DNA samples, thereby eliminating 6 markers from the list of candidates. The remaining candidate, the ERBB3 gene microsatellite, was genotyped in additional 245 MS patients and controls. No association of the ERBB3 marker with the disease was detected in these additional cohorts. In consequence, we did not find further evidence for apoptosis-related genes as predisposition factors in MS. [Abstract/Link to Full Text]

Carlsson E, Helgegren H, Slinde F
Resting energy expenditure is not influenced by classical music.
J Negat Results Biomed. 2005;46.
Obesity shows an increasing prevalence worldwide and a decrease in energy expenditure has been suggested to be one of the risk factors for developing obesity. An increase in resting energy expenditure would have a great impact on total energy expenditure. This study shows that classical music do not influence resting energy expenditure compared to complete silence. Further studies should be performed including other genres of music and other types of stress-inductors than music. [Abstract/Link to Full Text]

Laitala-Leinonen T
Unsatisfactory gene transfer into bone-resorbing osteoclasts with liposomal transfection systems.
J Negat Results Biomed. 2005;45.
BACKGROUND: Bone-resorbing osteoclasts are multinucleated cells that are formed via fusion of their hematopoietic stem cells. Many of the details of osteoclast formation, activation and motility remain unsolved. Therefore, there is an interest among bone biologists to transfect the terminally differentiated osteoclasts and follow their responses to the transgenes in vitro. Severe difficulties in transfecting the large, adherent osteoclasts have been encountered, however, making the use of modern cell biology tools in osteoclast research challenging. Transfection of mature osteoclasts by non-viral gene transfer systems has not been reported. RESULTS: We have systematically screened the usefulness of several commercial DNA transfection systems in human osteoclasts and their mononuclear precursor cell cultures, and compared transfection efficacy to adenoviral DNA transfection. None of the liposome-based or endosome disruption-inducing systems could induce EGFP-actin expression in terminally differentiated osteoclasts. Instead, a massive cell death by apoptosis was found with all concentrations and liposome/DNA-ratios tested. Best transfection efficiencies were obtained by adenoviral gene delivery. Marginal DNA transfection was obtained by just adding the DNA to the cell culture medium. When bone marrow-derived CD34-positive precursor cells were transfected, some GFP-expression was found at the latest 24 h after transfection. Large numbers of apoptotic cells were found and those cells that remained alive, failed to form osteoclasts when cultured in the presence of RANKL and M-CSF, key regulators of osteoclast formation. In comparison, adenoviral gene delivery resulted in the transfection of CD34-positive cells that remained GFP-positive for up to 5 days and allowed osteoclast formation. CONCLUSION: Osteoclasts and their precursors are sensitive to liposomal transfection systems, which induce osteoclast apoptosis. Gene transfer to mononuclear osteoclast precursors or differentiated osteoclasts was not possible with any of the commercial transfection systems tested. Osteoclasts are non-dividing, adherent cells that are difficult to grow as confluent cultures, which may explain problems with transfection reagents. Large numbers of alphavbeta3 integrin on the osteoclast surface allows adenovirus endocytosis and infection proceeds in dividing and non-dividing cells efficiently. Viral gene delivery is therefore currently the method of choice for osteoclast transfection. [Abstract/Link to Full Text]

Vrbova L, Mamdani M, Moineddin R, Jaakimainen L, Upshur RE
Does socioeconomic status affect mortality subsequent to hospital admission for community acquired pneumonia among older persons?
J Negat Results Biomed. 2005;44.
BACKGROUND: Low socioeconomic status has been associated with increased morbidity and mortality for various health conditions. The purpose of this study was twofold: to examine the mortality experience of older persons admitted to hospital with community acquired pneumonia and to test the hypothesis of whether an association exists between socioeconomic status and mortality subsequent to hospital admission for community-acquired pneumonia. METHODS: A population based retrospective cohort study was conducted including all older persons patients admitted to Ontario hospitals with community acquired pneumonia between April 1995 and March 2001. The main outcome measures were 30 day and 1 year mortality subsequent to hospital admission for community-acquired pneumonia. RESULTS: Socioeconomic status for each patient was imputed from median neighbourhood income. Multivariate analyses were undertaken to adjust for age, sex, co-morbid illness, hospital and physician characteristics. The study sample consisted of 60,457 people. Increasing age, male gender and high co-morbidity increased the risk for mortality at 30 days and one year. Female gender and having a family physician as attending physician reduced mortality risk.The adjusted odds of death after 30-days for the quintiles compared to the lowest income quintile (quintile 1) were 1.02 (95% CI: 0.95-1.09) for quintile 2, 1.04 (95% CI: 0.97-1.12) for quintile 3, 1.01 (95% CI: 0.94-1.08) for quintile 4 and 1.03 (95% CI: 0.96-1.12) for the highest income quintile (quintile 5). For 1 year mortality, compared to the lowest income quintile the adjusted odds ratios were 1.01 (95% CI: 0.96-1.06) for quintile 2, 0.99 (95% CI: 0.94-1.04) for quintile 3, 0.99 (95% CI: 0.93-1.05) for quintile 4 and 1.03 (95% CI: 0.97-1.10) for the highest income quintile. CONCLUSION: Socioeconomic status is not associated with mortality in the older persons from community-acquired pneumonia in Ontario, Canada. [Abstract/Link to Full Text]

Shafiq N, Malhotra S, Pandhi P, Grover A
The "Statinth" wonder of the world: a panacea for all illnesses or a bubble about to burst.
J Negat Results Biomed. 2005;43.
After the introduction of statins in the market as effective lipid lowering agents, they were shown to have effects other than lipid lowering. These actions were collectively referred to as 'pleiotropic actions of statins.' Pleiotropism of statins formed the basis for evaluating statins for several indications other than lipid lowering. Evidence both in favour and against is available for several of these indications. The current review attempts to critically summarise the available data for each of these indications. [Abstract/Link to Full Text]

Paul DR, Kramer M, Rhodes DG, Rumpler WV
Preprandial ghrelin is not affected by macronutrient intake, energy intake or energy expenditure.
J Negat Results Biomed. 2005;42.
BACKGROUND: Ghrelin, a peptide secreted by endocrine cells in the gastrointestinal tract, is a hormone purported to have a significant effect on food intake and energy balance in humans. The influence of factors related to energy balance on ghrelin, such as daily energy expenditure, energy intake, and macronutrient intake, have not been reported. Secondly, the effect of ghrelin on food intake has not been quantified under free-living conditions over a prolonged period of time. To investigate these effects, 12 men were provided with an ad libitum cafeteria-style diet for 16 weeks. The macronutrient composition of the diets were covertly modified with drinks containing 2.1 MJ of predominantly carbohydrate (Hi-CHO), protein (Hi-PRO), or fat (Hi-FAT). Total energy expenditure was measured for seven days on two separate occasions (doubly labeled water and physical activity logs). RESULTS: Preprandial ghrelin concentrations were not affected by macronutrient intake, energy expenditure or energy intake (all P > 0.05). In turn, daily energy intake was significantly influenced by energy expenditure, but not ghrelin. CONCLUSION: Preprandial ghrelin does not appear to be influenced by macronutrient composition, energy intake, or energy expenditure. Similarly, ghrelin does not appear to affect acute or chronic energy intake under free-living conditions. [Abstract/Link to Full Text]

Jagiello P, Wieczorek S, Yu P, Csernok E, Gross WL, Epplen JT
Association study with Wegener granulomatosis of the human phospholipase Cgamma2 gene.
J Negat Results Biomed. 2005;41.
BACKGROUND: Wegener Granulomatosis (WG) is a multifactorial disease of yet unknown aetiology characterized by granulomata of the respiratory tract and systemic necrotizing vasculitis. Analyses of candidate genes revealed several associations, e.g. with alpha(1)-antitrypsin, proteinase 3 and with the HLA-DPB1 locus. A mutation in the abnormal limb mutant 5 (ALI5) mouse in the region coding for the hydrophobic ridge loop 3 (HRL3) of the phospholipaseCgamma2 (PLCgamma-2) gene, corresponding to human PLCgamma-2 exon 27, leads to acute and chronic inflammation and granulomatosis. For that reason, we screened exons 11, 12 and 13 coding for the hydrophobic ridge loop 1 and 2 (HRL1 and 2, respectively) and exon 27 of the PLCgamma-2 protein by single strand conformation polymorphism (SSCP), sequencing and PCR/ restriction fragment length polymorphism (RFLP) analyses. In addition, we screened indirectly for disease association via 4 microsatellites with pooled DNA in the PLCgamma-2 gene. RESULTS: Although a few polymorphisms in these distinct exons were observed, significant differences in allele frequencies were not identified between WG patients and respective controls. In addition, the microsatellite analyses did not reveal a significant difference between our patient and control cohort. CONCLUSION: This report does not reveal any hints for an involvement of the PLCgamma-2 gene in the pathogenesis of WG in our case-control study. [Abstract/Link to Full Text]

Olsson M, English MA, Mason J, Licht JD, Ekblom P
Despite WT1 binding sites in the promoter region of human and mouse nucleoporin glycoprotein 210, WT1 does not influence expression of GP210.
J Negat Results Biomed. 2004;37.
BACKGROUND: Glycoprotein 210 (GP210) is a transmembrane component of the nuclear pore complex of metazoans, with a short carboxyterminus protruding towards the cytoplasm. Its function is unknown, but it is considered to be a major structural component of metazoan nuclear pores. Yet, our previous findings showed pronounced differences in expression levels in embryonic mouse tissues and cell lines. In order to identify factors regulating GP210, the genomic organization of human GP210 was analyzed in silico. RESULTS: The human gene was mapped to chromosome 3 and consists of 40 exons spread over 102 kb. The deduced 1887 amino acid showed a high degree of alignment homology to previously reported orthologues. Experimentally we defined two transcription initiation sites, 18 and 29 bp upstream of the ATG start codon. The promoter region is characterized by a CpG island and several consensus binding motifs for gene regulatory transcription factors, including clustered sites associated with Sp1 and the Wilms' tumor suppressor gene zinc finger protein (WT1). In addition, distal to the translation start we found a (GT)n repetitive sequence, an element known for its ability to bind WT1. Homologies for these motifs could be identified in the corresponding mouse genomic region. However, experimental tetracycline dependent induction of WT1 in SAOS osteosarcoma cells did not influence GP210 transcription. CONCLUSION: Although mouse GP210 was identified as an early response gene during induced metanephric kidney development, and WT1 binding sites were identified in the promoter region of the human GP210 gene, experimental modulation of WT1 expression did not influence expression of GP210. Therefore, WT1 is probably not regulating GP210 expression. Instead, we suggest that the identified Sp binding sites are involved. [Abstract/Link to Full Text]

Stenberg AE, Sylvén L, Magnusson CG, Hultcrantz M
Immunological parameters in girls with Turner syndrome.
J Negat Results Biomed. 2004;36.
Disturbances in the immune system has been described in Turner syndrome, with an association to low levels of IgG and IgM and decreased levels of T- and B-lymphocytes. Also different autoimmune diseases have been connected to Turner syndrome (45, X), thyroiditis being the most common.Besides the typical features of Turner syndrome (short stature, failure to enter puberty spontaneously and infertility due to ovarian insufficiency) ear problems are common (recurrent otitis media and progressive sensorineural hearing disorder).Levels of IgG, IgA, IgM, IgD and the four IgG subclasses as well as T- and B-lymphocyte subpopulations were investigated in 15 girls with Turners syndrome to examine whether an immunodeficiency may be the cause of their high incidence of otitis media. No major immunological deficiency was found that could explain the increased incidence of otitis media in the young Turner girls. [Abstract/Link to Full Text]

Tanner S, Barberis A
CP-31398, a putative p53-stabilizing molecule tested in mammalian cells and in yeast for its effects on p53 transcriptional activity.
J Negat Results Biomed. 2004;35.
BACKGROUND: CP-31398 is a small molecule that has been reported to stabilize the DNA-binding core domain of the human tumor suppressor protein p53 in vitro. The compound was also reported to function as a potential anti-cancer drug by rescuing the DNA-binding activity and, consequently, the transcription activation function of mutant p53 protein in mammalian tissue culture cells and in mice. RESULTS: We performed a series of gene expression experiments to test the activity of CP-31398 in yeast and in human cell cultures. With these cell-based assays, we were unable to detect any specific stimulation of mutant p53 activity by this compound. Concentrations of CP-31398 that were reported to be active in the published work were highly toxic to the human H1299 lung carcinoma and Saos-2 cell lines in our experiments. CONCLUSION: In our experiments, the small molecule CP-31398 was unable to reactivate mutant p53 protein. The results of our in vivo experiments are in agreement with the recently published biochemical analysis of CP-31398 showing that this molecule does not bind p53 as previously claimed, but intercalates into DNA. [Abstract/Link to Full Text]

Lim ET, Petzold A, Leary SM, Altmann DR, Keir G, Thompson EJ, Miller DH, Thompson AJ, Giovannoni G
Serum S100B in primary progressive multiple sclerosis patients treated with interferon-beta-1a.
J Negat Results Biomed. 2004;34.
S100B belongs to a family of calcium-binding proteins implicated in intracellular and extracellular regulatory activities. This study of serum S100B in primary progressive multiple sclerosis (PPMS) is based on data obtained from a randomized, controlled trial of Interferon beta-1a in subjects with PPMS. The key questions were whether S100B levels were associated with either disability or MRI findings in primary progressive MS and whether Interferon beta-1a has an effect on their S100B levels. Serial serum S100B levels were measured using an ELISA method. The results demonstrated that serum S100B is not related to either disease progression or MRI findings in subjects with primary progressive MS given Interferon beta-1a. Furthermore there is no correlation between S100B levels and the primary and secondary outcome measures. [Abstract/Link to Full Text]

Teixeira OU, Bortolotto LA, Silva HB
The contrast-enhanced Doppler ultrasound with perfluorocarbon exposed sonicated albumin does not improve the diagnosis of renal artery stenosis compared with angiography.
J Negat Results Biomed. 2004;33.
There are no studies investigating the effect of the contrast infusion on the sensitivity and specificity of the main Doppler criteria of renal artery stenosis (RAS). Our aim was to evaluate the accuracy of these Doppler criteria prior to and following the intravenous administration of perfluorocarbon exposed sonicated albumin (PESDA) in patients suspected of having RAS. Thirty consecutive hypertensive patients (13 males, mean age of 57 +/- 10 years) suspected of having RAS by clinical clues, were submitted to ultrasonography (US) of renal arteries before and after enhancement using continuous infusion of PESDA. All patients underwent angiography, and haemodynamically significant RAS was considered when >or=50%. At angiography, it was detected RAS >or=50% in 18 patients, 5 with bilateral stenosis. After contrast, the examination time was slightly reduced by approximately 20%. In non-enhanced US the sensitivity was better when based on resistance index (82.9%) while the specificity was better when based on renal aortic ratio (89.2%). The predictive positive value was stable for all indexes (74.0%-88.0%) while negative predictive value was low (44%-51%). The specificity and positive predictive value based on renal aortic ratio increased after PESDA injection respectively, from 89 to 97.3% and from 88 to 95%. In hypertensives suspected to have RAS the sensitivity and specificity of Duplex US is dependent of the criterion evaluated. Enhancement with continuous infusion of PESDA improves only the specificity based on renal aortic ratio but do not modify the sensitivity of any index. [Abstract/Link to Full Text]

Hanson DA, Ziegler SF
Fusion of green fluorescent protein to the C-terminus of granulysin alters its intracellular localization in comparison to the native molecule.
J Negat Results Biomed. 2004;32.
The engineering of green fluorescent protein (GFP) fusion constructs in order to visibly tag a protein of interest has become a commonly used cell biology technique. Although caveats to this approach are obvious, literature reports in which the chimeric molecule behaves differently than the native molecule are scant. This brief report describes one such case. Granulysin, a small lytic and antimicrobial protein produced by cytotoxic lymphocytes, traffics to the regulated secretory system and is subsequently released from cells upon proper stimulus. In an attempt to elucidate mechanisms by which it accumulates in and is released from cytolytic granules, GFP was fused to the C-terminus of granulysin and expressed in an NK cell line. A control construct expressing the native protein was similarly expressed. The data demonstrate that, while the fusion protein is expressed and secreted, its subcellular localization is altered in comparison to native granulysin. Thus, the addition of GFP to the C-terminus of granulysin obscures the signal(s) that cytotoxic lymphocytes use to sort it to the regulated secretory pathway despite its normal biosynthesis and secretion. This example is offered as a cautionary account for other researchers who contemplate using this technology. [Abstract/Link to Full Text]

Joos B, Uebelhart D, Michel BA, Sprott H
Influence of an outpatient multidisciplinary pain management program on the health-related quality of life and the physical fitness of chronic pain patients.
J Negat Results Biomed. 2004;31.
BACKGROUND: Approximately 10 to 20 percent of the population is suffering from chronic pain. Since this represents a major contribution to the costs of the health care system, more efficient measures and interventions to treat these patients are sought. RESULTS: The development of general health and physical activity of patients with chronic pain was assessed in an interdisciplinary outpatient pain management program (IOPP). 36 patients with an average age of 48 years were included in the IOPP. Subjective assessment of well-being was performed at five time points (baseline, post intervention and 3, 6, and 12 months thereafter) by using standardized questionnaires. The study focused on the quality of life survey Medical Outcomes Study Short Form-36, which is a validated instrument with established reliability and sensitivity. In addition, the patients participated in physical assessment testing strength, power, endurance, and mobility.Prior to therapy a substantial impairment was found on different levels. Marked improvements in the psychological parameters were obtained by the end of the program. No success was achieved with regard to the physical assessments. CONCLUSION: Although many different studies have evaluated similar programs, only few of them have attained positive results such as improvements of general quality of life or of physical strength. Often no difference from the control group could be detected only some months after the intervention. In the present study no significant persistent improvement of well-being occurred. Possible reasons are either wrong instruments, wrong selection of patients or wrong interventions. [Abstract/Link to Full Text]

Nöller V, Sprott H
Prospective epidemiological observations on the course of the disease in fibromyalgia patients.
J Negat Results Biomed. 2003;24.
OBJECTIVES: The aim of the study was to carry out a survey in patients with fibromyalgia (FM), to examine their general health status and work incapacity (disability-pension status), and their views on the effectiveness of therapy received, over a two-year observation period. METHODS: 48 patients diagnosed with FM, according to the American College of Rheumatology (ACR) criteria, took part in the study. At baseline, and on average two years later, the patients underwent clinical investigation (dolorimetry, laboratory diagnostics, medical history taking) and completed the Fibromyalgia questionnaire (Dettmer and Chrostek 1). RESULTS: 27/48 (56%) patients participated in the two-year follow-up. In general, the patients showed no improvement in their symptoms over the observation period, regardless of the type of therapy they had received. General satisfaction with quality of life improved, as did satisfaction regarding health status and the family situation, although the degree of pain experienced remain unchanged. In comparison with the initial examination, there was no change in either work-capacity or disability-pension status. CONCLUSIONS: The FM patients showed no improvement in pain, despite the many various treatments received over the two-year period. The increase in general satisfaction over the observation period was believed to be the result of patient instruction and education about the disease. To what extent a population of patients with FM would show similar outcomes if they did not receive any instruction/education about their disorder, cannot be ascertained from the present study; and, indeed, the undertaking of a study to investigate this would be ethically questionable. As present, no conclusions can be made regarding the influence of therapy on the primary and secondary costs associated with FM. [Abstract/Link to Full Text]

Ebenfelt A
Bacterial adherence to mucosal epithelium in the upper airways has less significance than believed.
J Negat Results Biomed. 2003;23.
BACKGROUND: Bacterial adherence to the upper airway epithelium is considered to be an important phenomenon in the pathogenesis of infections. However, the evidence for the hypothesis that bacterial adherence to mucosal epithelial cells has significance for pathogenesis of mucosal infections is based on studies using indirect techniques. We could find no biopsy studies with direct ocular observations of significant numbers of bacteria adhering to upper airway mucosal epithelial cells either in health or during disease. RESULTS: We studied specimens from healthy and infected tonsillar epithelium and specimens from the soft palate epithelium obtained by surgery. The specimens were examined by TEM. In the vast majority of specimens, we found no bacteria adhering to the epithelial cells in the mucosal line regardless of whether the patient was infected or not. Bacteria adhering to shed epithelial cells were seen in higher numbers. Furthermore, as bacteria are small compared to epithelial cells, we calculated the risk of overlooking every adhered bacteria in a section if bacterial adherence was such a significant phenomenon as earlier suggested. We found this risk to be very small. CONCLUSION: We conclude that bacterial adherence to mucosal surface epithelial cells is not a significant phenomenon, either in healthy mucosa in the upper airways or during infection. This is also in line with our earlier results, where we have shown that the site for the infectious process in pharyngotonsillitis is in the secretion on the tonsillar mucosal surface. [Abstract/Link to Full Text]

Jaspan HB, Richard Gaumer H, Garry RF
Quantitative competitive reverse transcription polymerase chain reaction is not a useful method for quantification of CD4 and CD8 cell status during HIV infection.
J Negat Results Biomed. 2003;22.
BACKGROUND: A polymerase chain reaction (PCR)-based method for quantitating CD4 and CD8 mRNA could provide a means of assessing immune status of AIDS patients and other immunologically compromised persons without requiring large blood draws, and could be exquisitely sensitive. Such a method would also be useful in assessing the immune status of patients retrospectively. RESULTS: Quantitative competitive reverse transcription PCR (QC-RT-PCR) assays were developed for measurement of CD4 and CD8 mRNA. Samples were obtained from HIV-positive and negative patients whose CD4 and CD8 counts had been determined via Flow Cytometry. The quantity of CD4 (n = 13) and CD8 (n = 28) mRNA standardized according to GAPDH mRNA quantities, all determined by QC-RT-PCR, were compared to cell number as determined by flow cytometry. There was no correlation between CD4 and CD8 cell counts and mRNA levels of CD4 and CD8 as determined by QC-RT-PCR. There is no correlation between CD4 and CD8 mRNA levels and the number of cells expressing these proteins on their surface. CONCLUSION: QC-RT-PCR, and related methodologies are not useful substitutes for assessment of CD4 and CD8 cell numbers in HIV-infected persons. [Abstract/Link to Full Text]

Hoke DE, Carson DD, Höök M
A heparin binding synthetic peptide from human HIP / RPL29 fails to specifically differentiate between anticoagulantly active and inactive species of heparin.
J Negat Results Biomed. 2003;21.
Despite extensive progress in determining structures within heparin and heparan sulfate (Hp/HS) and the discovery of numerous proteinaceous binding partners for Hp/HS so far; the only detailed characterization of a specific protein-glycosaminoglycan interaction is antithrombin III (ATIII) binding to a Hp pentasaccharide containing a unique 3-O-sulfated glucosamine residue. Previously, it was reported from our laboratories that a 16 amino acid synthetic peptide derived from the C-terminus of human HIP/RPL29 (HIP peptide-1) enriched for ATIII-dependent anticoagulant activity, presumably by specifically binding the ATIII pentasaccharide. Herein, we demonstrate that HIP peptide-1 cannot enrich ATIII-dependent anticoagulant activity from a starting pool of porcine intestinal mucosa Hp through a bio-specific interaction. However, a HIP peptide-1 column can be used to enrich for anticoagulantly active Hp from a diverse pool of glycosaminoglycans known as Hp byproducts by a mechanism of nonspecific charge interactions. Thus, HIP peptide-1 cannot recognize Hp via bio-specific interactions but binds glycosaminoglycans by non-specific charge interactions. [Abstract/Link to Full Text]

Pfeffer C, Olsen BR
Editorial: Journal of negative results in biomedicine.
J Negat Results Biomed. 2002;12. [Abstract/Link to Full Text]

Hebert RS, Wright SM, Dittus RS, Elasy TA
Prominent medical journals often provide insufficient information to assess the validity of studies with negative results.
J Negat Results Biomed. 2002;11.
BACKGROUND: Physicians reading the medical literature attempt to determine whether research studies are valid. However, articles with negative results may not provide sufficient information to allow physicians to properly assess validity. METHODS: We analyzed all original research articles with negative results published in 1997 in the weekly journals BMJ, JAMA, Lancet, and New England Journal of Medicine as well as those published in the 1997 and 1998 issues of the bimonthly Annals of Internal Medicine (N = 234). Our primary objective was to quantify the proportion of studies with negative results that comment on power and present confidence intervals. Secondary outcomes were to quantify the proportion of these studies with a specified effect size and a defined primary outcome. Stratified analyses by study design were also performed. RESULTS: Only 30% of the articles with negative results comment on power. The reporting of power (range: 15%-52%) and confidence intervals (range: 55-81%) varied significantly among journals. Observational studies of etiology/risk factors addressed power less frequently (15%, 95% CI, 8-21%) than did clinical trials (56%, 95% CI, 46-67%, p < 0.001). While 87% of articles with power calculations specified an effect size the authors sought to detect, a minority gave a rationale for the effect size. Only half of the studies with negative results clearly defined a primary outcome. CONCLUSION: Prominent medical journals often provide insufficient information to assess the validity of studies with negative results. [Abstract/Link to Full Text]

Recent Articles in The Canadian Journal of Clinical Pharmacology

Ipp M, Cohen E, Goldbach M, Macarthur C
Pain response to M-M-R vaccination in 4-6 year old children.
Can J Clin Pharmacol. 2006;13(3):e296-9.
BACKGROUND: Differences in pain response to two different M-M-R products have previously been demonstrated in 12-month old infants and in 4 â 6 year old children. Objective To determine if the acute and immediate pain response to two licensed M-M-R vaccine products (using a self-report measure) in children 4-6 years of age was similar to that demonstrated in younger infants. METHODS: Randomized, double blind, study. Subjects were randomly allocated to PriorixA (SmithKline Beecham) or M-M-R IIA (Merck Frosst). The primary outcome measure was pain response to vaccination quantified using a self-report OUCHER pain scale. Secondary outcome measures included pain measurement by proxy (physician and parent) using a visual analog scale (VAS) and measurement of cry and cry duration immediately post-vaccination. RESULTS: Of the 60 subjects enrolled, 30 received PriorixA and 30 received M-M-R IIA. There were no significant differences between the two groups on age, sex, or previous painful procedure. Post-vaccination, children in the M-M-R IIA group had higher median pain scores compared with children in the PriorixA group for VAS (12.5 vs. 2.0, respectively by paediatricians, p=0.017; 18.5 vs. 5.0, respectively by parents, p=0.235), OUCHER (20 vs. 0.00, respectively, p=0.047). The median duration of crying post M-M-R IIA was higher compared with PriorixA (6 vs. 0 seconds, respectively, p=0.020). Conclusion PriorixA was associated with significantly less pain compared with M-M-R IIA, at the time of injection. [Abstract/Link to Full Text]

Duggal J, Singh S, Kuchinic P, Butler P, Arora R
Utility of esmolol in thyroid crisis.
Can J Clin Pharmacol. 2006;13(3):e292-5.
Thyroid storm is an uncommon but potentially life-threatening manifestation of hyperthyroidism. Mortality can be 30-60% in hospitalized patients unless appropriately treated by combined therapy. We report a case of a 25-year-old African American woman with past medical history of Graves disease and moderately persistent asthma who presented to the emergency department with signs and symptoms of thyrotoxic crisis. Therapy instituted and included the use of an esmolol infusion for control of hypersympathetic activity. A review of the clinical presentation, diagnosis, and management of thyrotoxic crisis is presented along with a discussion on the choice of beta blockade therapy. [Abstract/Link to Full Text]

Kozer E, Scolnik D, Macpherson A, Rauchwerger D, Koren G
The effect of a short tutorial on the incidence of prescribing errors in pediatric emergency care.
Can J Clin Pharmacol. 2006;13(3):e285-91.
BACKGROUND: In the paediatric emergency department (ED) trainees are more likely to commit prescribing errors. OBJECTIVE: To determine whether a short educational intervention reduces the incidence of prescribing errors among trainees in a pediatric ED. METHODS: A prospective cohort study at the ED of a tertiary paediatric hospital. All fellows and residents arriving at the ED at the beginning of the academic year were invited to participate in a 30-minute tutorial focusing on appropriate methods for prescribing medications, followed by a written test. Eighteen days were selected randomly during July 2001. All the charts from these days were reviewed for medication errors. Two reviewers, blinded to whether or not a particular physician attended the tutorial, independently decided whether or not an error had occurred. The main outcome measure was the number of prescribing errors. RESULTS: Twenty-two trainees worked in the ED during July 2001. Of these, 13 trainees attended the tutorial. Eight hundred and ninety nine orders given by trainees were evaluated. We identified 66 (12.4%) errors in 533 orders given by those who attended tutorial, and 46 (12.7%) errors in 363 orders given by those who did not attend tutorial. The adjusted odds of a medication error was not significantly different between those who did not attend the tutorial and those who did (OR: 1.07 95% CI: 0.66-1.70). CONCLUSIONS: A short tutorial, followed by a written test, administered to trainees before entering their rotation in the paediatric ED, did not appear to reduce prescribing errors. [Abstract/Link to Full Text]

Dugoua JJ, Mills E, Perri D, Koren G
Safety and efficacy of ginkgo (Ginkgo biloba) during pregnancy and lactation.
Can J Clin Pharmacol. 2006;13(3):e277-84.
BACKGROUND: There is a lack of basic knowledge on the part of both clinicians and patients as to the indications for use and safety of herbal medicines used in pregnancy and lactation. This is one article in a series that systematically reviews the evidence for commonly used herbs during pregnancy and lactation. OBJECTIVES: To systematically review the literature for evidence on the use, safety, and pharmacology of ginkgo focusing on issues pertaining to pregnancy and lactation. METHODS: We searched 7 electronic databases and compiled data according to the grade of evidence found. RESULTS: There is some very weak scientific evidence from animal and in vitro studies that ginkgo leaf has antiplatelet activity, which may be of concern during labour as ginkgo use could prolong bleeding time. Low-level evidence based on expert opinion shows that ginkgo leaf may be an emmenagogue and have hormonal properties. The safety of ginkgo leaf during lactation is unknown. Patients and clinicians should be aware of past reports of ginkgo products being adulterated with colchicine. CONCLUSIONS: Ginkgo should be used with caution during pregnancy, particularly around labour where its anti-platelet properties could prolong bleeding time. During lactation the safety of ginkgo leaf is unknown and should be avoided until high quality human studies are conducted to prove its safety. [Abstract/Link to Full Text]

Dugoua JJ, Mills E, Perri D, Koren G
Safety and efficacy of St. John's wort (hypericum) during pregnancy and lactation.
Can J Clin Pharmacol. 2006;13(3):e268-76.
BACKGROUND: There is a lack of basic knowledge on the part of both clinicians and patients as to the indications for use and safety of herbal medicines in pregnancy and lactation. This is one article in a series that systematically reviews the evidence for commonly used herbs during pregnancy and lactation. OBJECTIVES: To systematically review the literature for evidence on the use, safety, and pharmacology of St. John's wort focusing on issues pertaining to pregnancy and lactation. METHODS: We searched 7 electronic databases and compiled data according to the grade of evidence found. RESULTS: There is very weak scientific evidence based on a case report that St Johns wort is of minimal risk when taken during pregnancy. There is in vitro evidence from animal studies that St John's wort during pregnancy does not affect cognitive development nor cause long-term behavioral defects, but may lower offspring birth weight. There is weak scientific evidence that St. John's wort use during lactation does not affect maternal milk production nor affect infant weight, but, in a few cases, may cause colic, drowsiness or lethargy. There is weak scientific evidence that St John's wort induces CYP450 enzymes, which may lower serum medication levels below therapeutic range; this may be of concern when administering medications during pregnancy and lactation. CONCLUSIONS: Caution is warranted with the use of St John's wort during pregnancy until further high quality human research is conducted to determine its safety. St John's wort use during lactation appears to be of minimal risk, but may cause side effects. Caution is warranted when using medications along with St John's wort. [Abstract/Link to Full Text]

Perri D, Dugoua JJ, Mills E, Koren G
Safety and efficacy of echinacea (Echinacea angustafolia, e. purpurea and e. pallida) during pregnancy and lactation.
Can J Clin Pharmacol. 2006;13(3):e262-7.
BACKGROUND: There is a lack of basic knowledge on the part of both clinicians and patients as to the indications for use and safety of herbal medicines used in pregnancy and lactation. This is one article in a series that systematically reviews the evidence for commonly used herbs during pregnancy and lactation. OBJECTIVES: To systematically review the literature for evidence on the use, safety, and pharmacology of echinacea focusing on issues pertaining to pregnancy and lactation. METHODS: We searched 7 electronic databases and compiled data according to the grade of evidence found. RESULTS: There is good scientific evidence from a prospective cohort study that oral consumption of echinacea during the first trimester does not increase the risk for major malformations. Low-level evidence based on expert opinion shows that oral consumption of echinacea in recommended doses is safe for use during pregnancy and lactation. CONCLUSIONS: Echinacea is non-teratogenic when used during pregnancy. Caution with using Echinacea during lactation until further high quality human studies can determine its safety. [Abstract/Link to Full Text]

Dugoua JJ, Seely D, Perri D, Koren G, Mills E
Safety and efficacy of black cohosh (Cimicifuga racemosa) during pregnancy and lactation.
Can J Clin Pharmacol. 2006;13(3):e257-61.
BACKGROUND: There is a lack of basic knowledge on the part of both clinicians and patients as to the indications for use and safety of herbal medicines in pregnancy and lactation. This is one article in a series that systematically reviews the evidence for commonly used herbs during pregnancy and lactation. OBJECTIVES: To systematically review the literature for evidence on the use, safety, and pharmacology of black cohosh focusing on issues pertaining to pregnancy and lactation. METHODS: We searched 7 electronic databases and compiled data according to the grade of evidence found. RESULTS: Black cohosh, alone or in combination with other medicinal herbs as "mother's cordial", has a long traditional use and is frequently used by midwives as a uterine stimulant and labour-inducing aid. Low-level evidence based on theory and expert opinion shows the following concerns with respect to black cohosh use during pregnancy: 1) labour-inducing effects, 2) hormonal effects, 3) emmenagogue properties and, 4) anovulatory effects. During lactation, there is low-level evidence from theoretical and expert opinion of black cohosh having hormonal properties. CONCLUSIONS: Black cohosh should be used with caution during pregnancy, particularly during the first trimester where its purported labour-inducing effects could be of concern, and during lactation. Black cohosh should undergo rigorous high quality human studies to determine its safety in pregnancy and lactation. [Abstract/Link to Full Text]

Koren G, Oren D, Rouleau M, Carmeli D, Matsui D
Comparison of verbal claims for natural health products made by health food stores staff versus pharmacists in Ontario, Canada.
Can J Clin Pharmacol. 2006;13(2):e251-6.
BACKGROUND: This study tested the hypothesis that while there are no written medical claims existing for many NHP, such claims are made verbally, giving a false impression that these are proven medical products. OBJECTIVE: To compare the number and type of verbal claims for NHP made by pharmacists to those made by health food stores personnel. METHODS: Randomly selected Canadian pharmacies selling NHP and health food stores were visited and the staff was asked to recommend natural health products for the treatment of hypertension. RESULTS: All health food stores (n=20) but only 4 out of 38 pharmacies (p< 0.001) recommended NHP for the treatment of hypertension. A majority of health food store staff (70%) stated that NHP are superior or equal to medicinal drugs in treating hypertension based on efficacy. CONCLUSION: Unlike pharmacy practice, verbal claims are common practice in health food stores, despite the lack of either written claims and/or proof of efficacy for most of them. These may be a very effective approach given that 30-40% of North American adults are functionally illiterate. These verbal claims are often inappropriate and not evidence-based. [Abstract/Link to Full Text]

Singh SR, Levine MA
Natural health product use in Canada: analysis of the National Population Health Survey.
Can J Clin Pharmacol. 2006;13(2):e240-50.
BACKGROUND: The use of natural health products (NHPs) in Western countries has increased dramatically over the past two decades. Although prevalence estimates have been published in the U.S. and elsewhere, little is known about the characteristics of persons who use NHPs. OBJECTIVES: To measure the prevalence of NHP use among adults in Canada, identify the most commonly used agents, and determine the socioeconomic, demographic, and health-related correlates of use. METHODS: NHP use by adults was assessed using the 2000-2001 National Population Health Survey (NPHS), a biennial general health survey conducted by Statistics Canada. A total of 11,424 adults completed the survey in 2000-2001. NHPs were defined as botanical and naturally-derived non-botanical products, excluding essential vitamins and minerals. Prevalence of use estimates were calculated nationally, and by age, gender, socioeconomic status, disease states, and health care practices. Multivariate logistic regression modeling was used to simultaneously assess the correlations of these variables with NHP use. RESULTS: The prevalence of past 2-day NHP use in Canada was 9.3% in 2000-2001. Fifty-seven percent of users also reported taking a conventional medicine in the same period. Glucosamine, echinacea, and garlic were the most frequently used products. Women reported NHP use more frequently than men (11.5% vs. 7.1%). As compared to young adults, NHP use was about 50% higher in middle-aged and older Canadians. There were no associations with either income or education level. Several disease states were associated with a high prevalence of NHP use: respondents with fibromyalgia (23.3%), inflammatory bowel disease (17.4%), and urinary incontinence (16.8%) were most likely to be NHP users. However, in the multivariate analysis, age and the use of vitamins or minerals were most predictive of NHP use, while health status variables were of less importance. CONCLUSIONS: NHP use is an important health phenomenon in Canada. Although respondents in poor health were more likely to use NHPs, a significant proportion of healthy Canadians also reported NHP use. The use of NHPs also cut across different socioeconomic groups. Concurrent use of conventional medications was common and suggests a need for health professionals to monitor for potential interactions. [Abstract/Link to Full Text]

Sedgeworth J, Derewlany L
Institution/investigator-initiated clinical trials in Canada.
Can J Clin Pharmacol. 2006;13(2):e236-9.
Institution/investigator-initiated clinical trials in Canada require adherence to similar regulations and processes required of large pharmaceutical manufacturers. This may require an investigator to file a Clinical Trial Application. This article outlines the responsibilities and actions that are required prior to, during, and after the conduct of an institution/investigator initiated trial. [Abstract/Link to Full Text]

Rezvani M, Hartfield D
Cocaine toxicity after laryngoscopy in an infant.
Can J Clin Pharmacol. 2006;13(2):e232-5.
Iatrogenic cocaine toxicity was observed in a 5.5-month-old male who received intranasal cocaine as a topical anesthetic for laryngoscopy. He became agitated, diaphoretic, tachycardic, and hypertensive shortly following the procedure. To control his signs and symptoms, he required 3 doses of IV lorazepam. Systemic absorption and toxicity can vary amongst individuals, making it difficult to determine appropriate dosing. The maximum dose of 1 mg/kg in children has not been validated and toxicity may appear at a much lower dose in certain individuals. Pediatric patients receiving topical cocaine as an anesthetic must be given the lowest possible dose, and then carefully monitored for signs of systemic absorption. [Abstract/Link to Full Text]

Baath NS, Hong J, Sattar SP
Possible carbamazepine toxicity with terbinafine.
Can J Clin Pharmacol. 2006;13(2):e228-31.
OBJECTIVE: To report a case of carbamazepine and terbinafine interaction resulting in an elevated level of carbamazepine and associated symptoms of toxicity. CASE SUMMARY: A 50-year-old Caucasian man developed an elevated carbamazapine level after starting terbinafine, which caused symptoms of toxicity with gait ataxia, dizziness and falls. DISCUSSION: This is a first report of a drug-drug interaction between carbamazapine and terbinafine. Although the mechanism for this interaction is not fully known, it is suspected that terbinafine decreased the metabolism of carbamazapine and led to increased levels that continued even days after the carbamazapine had been stopped. The Naranjo Scale suggests that this was a probable interaction (score 6). CONCLUSIONS: While several drug-drug interactions have been reported with carbamazapine, there are no previous reports of its interaction with terbinafine. Prescribers should exercise caution when prescribing these medications together. [Abstract/Link to Full Text]

Hemmelgarn B, Lévesque LE, Suissa S
Anti-diabetic drug use and the risk of motor vehicle crash in the elderly.
Can J Clin Pharmacol. 2006;13(1):e112-20.
BACKGROUND: Studies of the risk of motor vehicle crash associated with diabetes have produced conflicting results. OBJECTIVES: To assess whether the use of anti-diabetic drugs among the elderly increases the risk of motor vehicle crash. METHODS: The computerized databases of the various universal insurance programs of Québec were linked to form a cohort of all 224,734 elderly drivers that was followed from 1990-1993. Using a nested case-control approach, all 5,579 drivers involved in an injurious crash (cases) and a random sample of 13,300 control subjects were identified. Exposure to anti-diabetic drugs was assessed in the year preceding the index date, namely the date of the crash for the cases and a randomly selected date during follow-up for the controls. RESULTS: The adjusted rate ratio of an injurious crash was 1.4 (95% CI: 1.0-2.0) for current users of insulin monotherapy relative to non-users and 1.3 (95% CI: 1.0-1.7) for sulfonylurea and metformin combined. Monotherapy, using either a sulfonylurea or metformin, was not associated with an increased risk. There was a dose-response effect in subjects using high doses of combined oral therapy (RR 1.4; 95% CI: 1.0-2.0). For users of insulin monotherapy or of high doses of combined oral therapy, the increase corresponds to an excess rate of 32 crashes per 10,000 elderly drivers per year. CONCLUSIONS:L Elderly drivers treated with insulin monotherapy or a combination of sulfonylurea and metformin, especially at high doses, have a small increased risk of injurious crashes. There is no increased risk associated with any regimen of oral monotherapy. [Abstract/Link to Full Text]

Carrie AG, Kozyrskyj AL
Outpatient treatment of community-acquired pneumonia: evolving trends and a focus on fluoroquinolones.
Can J Clin Pharmacol. 2006;13(1):e102-11.
BACKGROUND: Increasing use of broad-spectrum antibiotics in the community, including fluoroquinolones, has been reported, despite concerns for developing antibiotic resistant organisms. Community-acquired pneumonia (CAP) is commonly treated on an outpatient basis, and recent treatment guidelines suggest only a limited role for fluoroquinolones. OBJECTIVES: To identify evolving trends in the outpatient treatment of CAP in adults, and to identify factors associated with receipt of a fluoroquinolone. METHODS: Retrospective observational design using population-based administrative data. Initial outpatient treatment for subjects diagnosed with CAP between May 1996 and March 2002 was examined. Logistic regression was used to examine the influence of patient characteristics on the receipt of a fluoroquinolone. RESULTS: A total of 31,940 outpatients with CAP were identified. The proportion of patients receiving fluoroquinolones increased from 6.6% in 1996/97 to 25.2% in 2001/02. Over the course of the study, 158 (25.9%) of the 610 patients meeting the eligibility criteria for treatment with fluoroquinolones, according to treatment guidelines, received these agents. Of the 31,330 subjects who did not meet the eligibility criterion, 3,886 (12.4%) received a fluoroquinolone. Other variables that influenced the receipt of a fluoroquinolone included: age (for every 10-year increase) [OR=1.16 (1.14-1.19)], urban residence [OR=1.40 (1.30-1.51)], presentation to an emergency department [OR=0.80 (0.70-0.90)], high-level drug use (six or more different drugs in the previous year) [OR=1.50 (1.41-1.59)], and income-level (highest to lowest) [OR=1.20 (1.08-1.35)]. CONCLUSION: The use of fluoroquinolones for the treatment of CAP is increasing. However less than 4% of the subjects receiving fluoroquinolones met eligibility requirements according to treatment guidelines. Initiatives to increase the uptake of treatment guidelines appear warranted. [Abstract/Link to Full Text]

Montori VM, Leung TW, Devereaux PJ, Schünemann HJ, Akl EA, Gafni A, Guyatt GH
Can contraindications compromise evidence-based, patient-centered clinical practice?
Can J Clin Pharmacol. 2006;13(1):e92-101.
BACKGROUND: Despite their often weak evidence base, contraindications convey the unequivocally adverse risk-benefit profile of an intervention in a specific clinical context. However, some patients in that context may nonetheless prefer the contraindicated intervention (with its potential benefits and risks) to the available alternatives. The impact of contraindications on treatment decisions remains unexplored. OBJECTIVE: To provide an estimate of the impact of the "contraindication" label on treatment decisions. METHODS: We conducted an international 6-wave email/internet and fax survey of practicing clinicians who were members of the American Diabetes Association or the College of Physicians and Surgeons of Ontario and had available email addresses and fax numbers. Each participant considered one of two patient scenarios. In each scenario, the patient expressed a strong preference for use of a medication that carried a "contraindication" label despite weak evidence of harm. We designed these scenarios so that respondents who placed greater weight on patient preferences and research evidence than on the label "contraindication" would be ready to prescribe the contraindicated medication. We determined the frequency with which the label "contraindication" dominated participants' treatment decisions despite patient preferences and weak evidence of harm. RESULTS: 466 participants responded (22% response rate). Depending on the group and scenario, contraindications dominated the decisions of 47% to 89% of surveyed clinicians, superseding patient preferences and research evidence. CONCLUSIONS: The label "contraindication" may often dominate clinicians' decisions about treatment and may compromise evidence-based, patient-centered clinical practice. Further research should elucidate the process that leads to the formulation of contraindications and its impact on treatment decision-making. [Abstract/Link to Full Text]

Mather CM
Medical innovation, unmet medical needs, and the drug pipeline.
Can J Clin Pharmacol. 2006;13(1):e85-91.
This paper outlines and illustrates the working of a theoretical approach from the social sciences for analyzing medical innovation, unmet medical need, and the drug pipeline. Using the social history of three drugs made from recombinant DNA (insulin, human growth hormone, and tissue-plasminogen activator) the paper shows how drugs can be both technically and organizationally efficient while the needs they satisfy can be created or identified. The paper posits that drugs that require more organizational efficiency tend to satisfy identified, rather then created needs. Key words: Recombinant DNA, technical efficiency, organizational efficiency, anthropology. [Abstract/Link to Full Text]

Gray J
Changing physician prescribing behaviour.
Can J Clin Pharmacol. 2006;13(1):e81-4.
Didactic approaches to educating physicians and/or other health professionals do not produce changes in learner behaviour. Similarly, printed materials and practice guidelines have not been shown to change prescribing behaviour. Evidence-based educational approaches that do have an impact on provider behaviour include: teaching aimed at identified learning needs; interactive educational activities; sequenced and multifaceted interventions; enabling tools such as patient education programs, flow charts, and reminders; educational outreach or academic detailing; and audit and feedback to prescribers. Dr. Jean Gray reflects over the past 25 years on how there has been a transformation in the types of activities employed to improve prescribing practices in Nova Scotia. The evolution of Continuing Medical Education (CME) has resulted in the creation of the Drug Evaluation Alliance of Nova Scotia (DEANS) program, which is one exemplar of an evidence-based educational approach to improving physician prescribing in that province. Key words: Evidence-based, education, prescribing. [Abstract/Link to Full Text]

Delaney JA, Lévesque LE, Etminan M, Suissa S
Furosemide use and hospitalization for benign prostatic hyperplasia.
Can J Clin Pharmacol. 2006;13(1):e75-80.
OBJECTIVE: Recent studies have shown that furosemide may have anti-inflammatory properties. We explored whether exposure to furosemide would reduce the risk of being hospitalized with prostatism, a marker of benign prostatic hyperplasia. METHODS: Using record linkage and the computerized health insurance databases of the province of Québec, Canada, we identified a cohort of men 65 years of age and older within which we conducted a case-control study. Cases were individuals hospitalized with prostatism (ICD-9 code 600) between January 1991 and June 1993, with the index date taken as the date of hospitalisation. Controls were those not having experienced the event during the study period, with an index date selected randomly during their follow-up. Cases and controls were required to have at least 2 (1/2) years of health coverage prior to index date in order to identify risk factors for benign prostatic hyperplasia and establish baseline medical history. We assessed the subjects' exposure to furosemide and various other diuretics in the period 180 to 900 days preceding the index date. Logistic regression was used to evaluate the association between the use of furosemide and hospitalization for prostatism, adjusting for potential confounders. RESULTS: The cohort included 8,814 subjects, of which 231 were cases and 8,583 controls. The rate of hospitalization for prostatism was lower for users of furosemide compared to non-users (adjusted rate ratio 0.49; 95% CI: 0.25-0.95). There was no association with the use of thiazide or potassium sparing diuretics (adjusted rate ratio 0.95; 95% CI: 0.65-1.37). Results suggestive of a protective effect associated with corticosteroid use were observed (adjusted rate ratio 0.64; 95% CI: 0.44-0.93). CONCLUSIONS: This study supports the hypothesis that furosemide can reduce the risk of hospitalization for prostatism, a marker of benign prostatic hyperplasia. [Abstract/Link to Full Text]

Bhanji NH, Chouinard G, Kolivakis T, Margolese HC
Persistent tardive rebound panic disorder, rebound anxiety and insomnia following paroxetine withdrawal: a review of rebound-withdrawal phenomena.
Can J Clin Pharmacol. 2006;13(1):e69-74.
OBJECTIVE: To describe tardive rebound anxiety phenomena (panic, anxiety and insomnia) following abrupt paroxetine discontinuation. METHOD: Case report, with comprehensive literature review on rebound and withdrawal phenomena associated with psychotropic medications. RESULTS: Three different discontinuation syndromes with psychotropics are described: (1) new-onset CNS-depressant type withdrawal symptoms (minor and major); (2) rebound syndromes; and (3) supersensitivity symptoms. Abrupt paroxetine discontinuation has been well described and fits the first category. Tardive rebound panic disorder-phenomena with paroxetine has some features of the supersensitivity category. CONCLUSION: Chronic paroxetine treatment may lead to 5-HT2-receptor down regulation, with desensitization of 5-HT1A and 5-HT2 receptors, which may contribute to tardive rebound symptoms upon abrupt withdrawal. Early reports suggest that genetic factors may also contribute to withdrawal symptoms in susceptible individuals. Cholinergic rebound may also occur and could explain tardive insomnia and anxiety in paroxetine withdrawal. [Abstract/Link to Full Text]

Campbell NR
The Canadian Hypertension Education Program (CHEP)--a therapeutic knowledge translation program.
Can J Clin Pharmacol. 2006;13(1):e65-8.
The Canadian Hypertension Education Program (CHEP) is a dynamic therapeutic knowledge translation program that changes annually based on the previous year's experience. To meet the challenge of hypertension treatment and control, CHEP activities include updating evidence-based management recommendations, implementing the recommendations and examining the impact of CHEP on hypertension management and hypertensive complications. CHEP aids health care professionals by providing credible, widely dissentinated up-to-date recommendations in multiple formats to suit individual learning needs. Key words: Hypertension, knowledge translation. [Abstract/Link to Full Text]

Shea SE
A comparison of methylphenidate formulations in the treatment of ADHD.
Can J Clin Pharmacol. 2006;13(1):e63-4. [Abstract/Link to Full Text]

Steele M, Weiss M, Swanson J, Wang J, Prinzo RS, Binder CE
A randomized, controlled effectiveness trial of OROS-methylphenidate compared to usual care with immediate-release methylphenidate in attention deficit-hyperactivity disorder.
Can J Clin Pharmacol. 2006;13(1):e50-62.
BACKGROUND: The thrice daily dosing regimen of immediate release methylphenidate (IR-MPH) for Attention Deficit/Hyperactivity Disorder (ADHD) requires in-school dosing, leading to issues surrounding dispensing and storage of controlled substances by school personnel and concerns over children?s privacy and the embarrassment associated with taking medication in public at school. OROS-methylphenidate (OROS-MPH) is a once-daily controlled-release formulation of methylphenidate (MPH) developed to overcome some of the limitations associated with IR-MPH and first-generation sustained-release formulations. Randomized, controlled trials (RCTs) that focus on treatment efficacy provide the best evidence for demonstrating whether an intervention works, but under ideal conditions one cannot discount the importance of efficacy study results. However, the most useful information to clinicians comes from an effectiveness study design. OBJECTIVES: To evaluate the effectiveness and tolerability of OROS-MPH versus usual care with IR-MPH in children aged 6 to 12 years with ADHD. METHODS: This 8 week, multicentre, open-label study randomized 147 subjects to either once-daily OROS-MPH or usual care with IR-MPH. Subjects were titrated to a clinically effective dose of either study medication over 4 weeks and maintained on that dose for an additional 4 weeks. The SNAP-IV parent-rating scale was used to assess effectiveness. RESULTS: OROS-MPH showed statistically significant superiority to IR-MPH in remission rate based on the 18 ADHD symptoms (p=0.0002, X2=13.8, df=1) and severity of ADHD and ODD symptoms (p=0.004, F=8.4, df=1,127), as well as on the following secondary assessments: IOWA Conners, Conners Parent Rating Scale (short version), Parent Stress Index, (short version); Visual Analogue Scale for social play; Clinical Global Impression-Severity, Clinical Global Impression-Improvement and Parent Satisfaction with treatment. OROS-MPH and IR-MPH were both well tolerated with a similar side effect profile. CONCLUSIONS: Once-daily OROS-MPH is significantly more effective than usual care with IR-MPH based on multiple outcome measures including remission rate. [Abstract/Link to Full Text]

Drug innovation and patient safety--the need for a new paradigm.
Can J Clin Pharmacol. 2006;13(1):e1-49. [Abstract/Link to Full Text]

van Mosseveld C
Pharmaceutical expenditure compared across countries.
Can J Clin Pharmacol. 2005;12(3):e269-75.
Pharmaceutical expenditures for 20 member countries of the Organisation for Economic Co-Operation and Development (OECD) are discussed and the appropriateness of making comparisons amongst these countries is addressed. This is a summary of a lecture presented at the International Conference on Pharmacoepidemiology in 2004. [Abstract/Link to Full Text]

Khaldi N, Miras A, Gromb S
Toxic epidermal necrolysis and clarithromycin.
Can J Clin Pharmacol. 2005;12(3):e264-8.
Toxic epidermal necrolysis almost always occurs after taking a medication. Despite spectacular clinical signs, it is mainly diagnosed with pathologic techniques. The identification of a drug as the cause for the immune related cytotoxic reaction can be difficult if the molecule is not generally known to be a classical cause of this reaction. The present study describes a female patient who rapidly developed a severe bullous skin disease after taking clarithromycin for tonsillitis. The case illustrates the process involved in attributing causality to a molecule using an established imputability assessment framework. [Abstract/Link to Full Text]

Jayaraman S, Rieder MJ, Matsui DM
Compliance assessment in drug trials: has there been improvement in two decades?
Can J Clin Pharmacol. 2005;12(3):e251-3.
BACKGROUND: Compliance is a key element in the success of therapy, both in practice and research. A study from 1974 demonstrated that compliance in clinical trials was only determined in 19% of studies requiring it. OBJECTIVES: The objective was to determine if there has been an improvement in compliance assessment in clinical trials. METHODS: All drug studies published in the British Medical Journal, Journal of Pediatrics, and Lancet from 1997 to 1999 were reviewed. Clinical trials were evaluated as to their measurement of compliance and the method of assessment. RESULTS: Of 303 studies in which the effects of drugs were reported, 165 required the incorporation of a measure of compliance, 86 did not, and in 52, compliance could not be measured. Of the studies requiring estimation of compliance, compliance was evaluated in 78 (47%). This rate did not vary between the journals examined or between trials in adults or children. The most common methods used to evaluate compliance were pill count (33%) and self report (25%). The use of drug assays (14%) and close supervision (9%) was less common. Electronic devices and other methods were uncommonly used (5%). In 16% of cases, a combination of methods was used. CONCLUSION: Although the rate of evaluation of compliance in drug trials has improved over the past 25 years, it continues to be examined in less than half of the clinical studies of drug effects in which compliance assessment is required. This rate appears to be similar in paediatric and adult drug studies. [Abstract/Link to Full Text]

Farahani P, Levine M, Gaebel K, Thabane L
Clinical data gap between phase III clinical trials (pre-marketing) and phase IV (post-marketing) studies: evaluation of etanercept in rheumatoid arthritis.
Can J Clin Pharmacol. 2005;12(3):e254-63.
BACKGROUND: There are fundamental differences in design between phase III clinical trials and phase IV post-marketing studies that involve patient characteristics, the clinical setting (environment) and the manner of drug use. As well, many phase IV studies are extensions of randomized clinical trials (RCTs) and suffer from selection bias. OBJECTIVE: To determine if the data obtained from RCTs of etanercept (Enbrel) in the treatment of rheumatoid arthritis would be representative of the effects attainable in community practice. METHOD: An analysis was conducted comparing data from published RCTs of etanercept use in rheumatoid arthritis patients with data collected in a community based cohort study that was not an extension of an RCT. RESULTS: Baseline clinical data, such as tender or painful joint count, patient's global assessment, the heath assessment questionnaire, physical and mental component summary of the SF-36, and rheumatoid arthritis drug profile were significantly different between the patients receiving etanercept in the phase IV community cohort study and the patients enrolled in the RCTs. Differences in the baseline data for the control patients were also noted amongst the RCT studies. The treatment outcome, American College of Rheumatology (ACR) response rate of 20%, 50% and 70% at 6 month, was the same between the cohort study and the RCTs, but at 12 months the clinical response was less for the community based patients than for the RCT patients. At 6 months there were fewer withdrawals involving community-based patients than RCT patients due to less frequent withdrawals associated with lack of efficacy. At 12 months the withdrawal rate due to either a lack of efficacy or from adverse events was similar between data sets. CONCLUSION: The data from the etanercept phase III RCTs may not reflect the characteristics of patients using etanercept in community practice, nor the clinical outcomes observed by RA patients at 12 months. These discrepancies may be derived from methodological differences in study design and patient selection. On the other hand, outcomes such as withdrawal rates at 12 months appear comparable between the two types of populations. [Abstract/Link to Full Text]

Levsky ME, Miller MA
Cardiovascular collapse from low dose bupivacaine.
Can J Clin Pharmacol. 2005;12(3):e240-5.
Bupivacaine is a long-acting local anesthetic agent that is very widely used for cutaneous infiltration, peripheral nerve blocks, epidural anesthesia, and spinal anesthesia. Well-described cardiotoxic effects of bupivacaine and other members of its class include dysrhythmias, hypotension, and depression of cardiac output. The minimum IV dose of bupivacaine previously associated with significant toxicity in humans is 1.6 mg/kg. A case is reported of bradyasystolic arrest with post resuscitation shock, without significant central nervous system (CNS) toxicity, following the injection of less than 1.1 mg/kg bupivacaine in an adult. [Abstract/Link to Full Text]

Pereira JA, Holbrook AM, Dolovich L, Goldsmith C, Thabane L, Douketis JD, Crowther M, Bates SM, Ginsberg JS
Are brand-name and generic warfarin interchangeable? A survey of Ontario patients and physicians.
Can J Clin Pharmacol. 2005;12(3):e229-39.
BACKGROUND: The issue of therapeutic equivalence has been a source of controversy in Canada since the approval of generic warfarin products in 2000. OBJECTIVES: We surveyed Ontario patients and physicians on perceptions of generic warfarin and brand substitution. METHODS: Self-administered questionnaires employed 7.0-point Likert scales of agreement. Patient participants were drawn from a thromboembolism clinic in Hamilton, Ontario. Physician participants were from a random sample of 375 Ontario family physicians, internists, cardiologists and hematologists. RESULTS: Eighty-one patients responded: 52% female, mean age 63.4 years and 63% brand-name warfarin users. Overall, 33% of respondents agreed or strongly agreed that they would feel comfortable taking generic warfarin. However, seventeen percent agreed or strongly agreed that generic warfarin was neither as safe nor as effective as brand-name warfarin, with this view more common amongst patients taking brand-name than those taking generic warfarin. One hundred and ten (29.3%) physicians returned the survey--29% females, mean age 45.3 years, 22% family physicians. Forty-four percent agreed or strongly agreed that they would rather prescribe brand-name than generic warfarin for patients starting warfarin therapy, while 40.7% agreed or strongly agreed that they would not feel comfortable switching from brand-name to generic warfarin. However, only 19.4% of physicians who had switched patients from brand-name to generic warfarin actually reported difficulties in managing the switch. CONCLUSION: While most patients and physicians appear to have accepted the principle of therapeutic equivalence of generic and brand-name warfarin, a sizable minority has concerns that could influence prescribing and compliance. [Abstract/Link to Full Text]

Lau L, Baruchel S
Can Canada sustain paediatric phase I trials? A national survey of cancer relapse in children.
Can J Clin Pharmacol. 2005;12(3):e222-8.
BACKGROUND: Paediatric phase I trials are critical to the evaluation of new agents using standardized methodology. However a large proportion of paediatric patients in Canada do not have access to phase I therapy. OBJECTIVES: A National Paediatric Cancer Relapse Survey was conducted to collect preliminary data to evaluate the feasibility of multi-centre paediatric phase I trials within Canada. METHODS: A survey consisting of 20 individual questions was sent out to all of the 17 paediatric oncology centres in Canada. RESULTS: Fifteen centres (88%) responded to the survey. 1027 children are diagnosed with cancer each year in Canada while 241 present with recurrent cancer. Of the 85 patients who are considered to be eligible for phase I study each year, only 53% were referred for phase I therapy. Two centres have more than 10 eligible patients a year, while the remaining 13 centres have less than 10 eligible patients each year. CONCLUSIONS: We estimate that 20% of the eligible patients could be accrued to phase I trials and Canada may provide sufficient patient number, i.e. 25 to 30 solid tumour patients every 2 years, to allow one multi-centre paediatric phase I trial to be completed over a 2-year period. [Abstract/Link to Full Text]

Recent Articles in Journal of Pharmacy & Pharmaceutical Sciences

Zarghi A, Rao PN, Knaus EE
Design and synthesis of new rofecoxib analogs as selective cyclooxygenase-2 (COX-2) inhibitors: replacement of the methanesulfonyl pharmacophore by a N-acetylsulfonamido bioisostere.
J Pharm Pharm Sci. 2007;10(2):159-67.
PURPOSE: A group of 3,4-diaryl- 2(5H)furanones were synthesized to determine whether a N-acetylsulfonamido (SO2NHCOCH3) moiety could be used as a bioisosteric replacement for the traditional sulfonamide (SO2NH2) and methanesulfonyl (SO2CH3) COX-2 pharmacophores. METHODS: In vitro COX-1 and COX-2 isozyme inhibition studies were carried out to acquire structure activity relationship data with respect to the point of attachment of the Nacetylsulfonamide moiety at the para and metapositions of the C-4 phenyl ring in conjunction with a variety of substituents (H, F, Cl, Me, OMe) at the para position of the C-3 phenyl ring. RESULTS: COX-1 and COX-2 inhibition studies showed that all compounds were selective inhibitors of COX-2 since no inhibition of COX-1 was observed at a concentration of 100 microM. The relative COX-2 potency, and COX-2 selectivity index, profiles for the C-4 para acetamidophenyl compounds, with respect to the C-3 phenyl parasubstituent was H > F > Cl. The point of attachment of the SO2NHCOCH3 substituent on the C-4 phenyl ring was a determinant of COX-2 potency, and COX-2 selectivity, where the relative activity profile was para acetylsulfonamido > meta acetylsulfonamido. 4-[4-(NAcetylsulfonamido) phenyl]-3-phenyl-2(5H)furanone was identified as a more potent (IC50 = 0.32 microM), and selective (S.I. > 313), COX-2 inhibitor than the parent reference compound rofecoxib (IC50 = 0.43 microM, S.I. > 232). CONCLUSIONS: The SO2NHCOCH3 moiety i) is a novel COX-2 pharmacophore that also has the potential to serve as a prodrug moiety to the traditional SO2NH2 COX-2 pharmacophore, and ii) it could serve as a useful COX-2 pharmacophore to study the structure-function relationship of the COX-2 isozyme in view of its potential to acetylate the NH2 moiety of amino acid residues such as Gln192 or Arg513 that line the pocket of the secondary COX-2 binding site. [Abstract/Link to Full Text]

Rajabalian S, Foroumadi A, Shafiee A, Emami S
Functionalized N(2-oxyiminoethyl) piperazinyl quinolones as new cytotoxic agents.
J Pharm Pharm Sci. 2007;10(2):153-8.
PURPOSE: The prokaryotic type II topoisomerases (DNA gyrase and topoisomerase IV) and the eukaryotic type II topoisomerases represent the cellular targets for quinolone antibacterial agents and a wide variety of anticancer drugs, respectively. In view of the mechanistic similarities and sequence homologies exhibited by the two enzymes, tentative efforts to selectively shift from an antibacterial to an antitumoral activity was made by synthesizing a series of functionalized N-(2-oxyiminoethyl)piperazinyl quinolones, in which the C-7 piperazine ring of antibacterial quinolones, ciprofloxacin and norfloxacin, is attached by a certain N-[2-(furan-2-yl)-2-oxyiminoethyl] and N-[2-(thiophen-2-yl)-2-oxyiminoethyl] moieties. Thus, as part of a continuing search for potential anticancer drug candidates in the N-substituted piperazinyl quinolones series, the cytotoxicity evaluation of functionalized N-(2-oxyiminoethyl) piperazinyl quinolones was our interest. METHODS: The growth inhibitory activities of synthesized N-[2-(furan-2-yl)-2-oxyiminoethyl] and N-[2-(thiophen-2-yl)-2-oxyiminoethyl] piperazinyl quinolones were determined against seven cancer cell lines using an in vitro cell culture system (MTT assay). RESULTS: Preliminary screening showed that some of N-(2-oxyiminoethyl) piperazinyl quinolone analogs containing O-benzyl group displayed in vitro cytotoxic activity comparable or higher than reference drug etoposide. CONCLUSIONS: These studies demonstrate that introduction of O-benzylmoiety on oxime group of N-(2-oxyimino) piperazinyl quinolone series changes the biological profile of piperazinyl quinolones from antibacterial to cytotoxic activity. As can be deduced from these data, O-benzyl functionalized N-(2-oxyiminoethyl) piperazinyl quinolones have excellent potential as a new class of cytotoxic agents. [Abstract/Link to Full Text]

Berger M, Kreutz FT, Horst JL, Baldi AC, Koff WJ
Phase I study with an autologous tumor cell vaccine for locally advanced or metastatic prostate cancer.
J Pharm Pharm Sci. 2007;10(2):144-52.
PURPOSE: To attempt the isolation and primary culture of prostate tumor cells, to use the cultured cells for active immunotherapy, and to evaluate the safety and efficacy in a Phase I clinical trial. MATERIALS AND METHODS: Tumor fragments were collected from 50 patients with prostate-specific antigen (PSA) > or = 10 ng/mL, < or = cT2 PCa who underwent radical retropubic prostatectomy (RRP) and 6 patients with metastatic PCa who underwent transurethral resection of the prostate (TURP). Cultured tumor cells were incubated with IFN-fi, irradiated, and cryopreserved. Seven vaccine inoculations were performed into > or = pT3 and/or N+ patients, and M+ patients, with the first two doses admixed with Bacille Calmette-Guerin (BCG). Follow-up was performed with measurement of delayed-type hypersensitivity (DTH) reactions, PSA and hemato-chemical tests, and bone scans. RESULTS: No cell culture was obtained in the TURP group. Cell culture and vaccine production were obtained in 37 cases (74%) in the RRP group. Eleven > or = pT3 and/or N+ patients were vaccinated. Toxicity was generally limited to the inoculation sites. DTH reactions > or = 10 mm were observed in 2 patients and > or = 5 mm in 6 patients. Two patients had a decrease in PSA levels after vaccine administration. CONCLUSIONS: The autologous cell vaccine is safe and seems to induce a positive immune cellular response. Primary cell culture and vaccine production can be obtained for most RRP patients, but not for TURP patients using our method. There seems to be some influence of the vaccine in PSA evolution after RRP. [Abstract/Link to Full Text]

Amsden B
Review of osmotic pressure driven release of proteins from monolithic devices.
J Pharm Pharm Sci. 2007;10(2):129-43.
Protein therapeutics are a rapidly growing drug class, with sales in 2004 in the area of $US 34 billion. They are presently administered primarily by injection, although there is increasing recognition that many proteins would benefit from long-term, localized delivery. Such delivery represents a significant challenge due principally to protein stability concerns. Polymeric delivery systems which rely on osmotic pressure driven drug release may prove to be an effective formulation approach. This paper reviews the evolution of osmotic pressure drug release from polymers, with an emphasis on their potential for protein delivery. It is concluded that osmotic pressure driven release is promising for protein delivery, but there is still a need for in vivo demonstration of protein stability and delivery efficacy. [Abstract/Link to Full Text]

Jain A, Gupta Y, Jain SK
Perspectives of biodegradable natural polysaccharides for site-specific drug delivery to the colon.
J Pharm Pharm Sci. 2007;10(1):86-128.
The ability to deliver drugs to the human colon in a specific manner has become feasible over the years. Targeting pharmaceutical drugs to the colon makes it possible to achieve local or systemic drug delivery to this site. To deliver the compounds in a non-degraded form to the lower part of the gastrointestinal tract, they must first of all pass through the stomach, the upper part of the intestine and must use the characteristics of the colon to specifically release the drugs in this part of the digestive tract. Among the different approaches available to achieve targeted drug release to the colon, the use of especially biodegradable polymers holds great promise. This family of natural polymers has great appeal to drug delivery as it is comprised of polymers with a large number of derivatizable groups, a wide range of molecular weights, varying chemical compositions, and, for the most part, low toxicity and biodegradability yet high stability. The most favorable property of these materials is their approval as pharmaceutical excipients. Polysaccharidases are bacterial enzymes that are available in sufficient quantity to be exploited in colon targeting. Bacterial sources of polysaccharidases as well as a detailed treatise of the enzymatic flora of the colonic region is reviewed, followed by a wide range of the polysaccharides which can be used solely for the purpose of colon-specific drug delivery. Present article also discusses few of the mechanisms by which members of the intestinal microbiota degrade complex polysaccharides. A final overview of the various approaches to target drugs to the colon utilizing natural polysaccharides, the limits and the future developments in this field with these natural polymers is discussed which leads to the conclusion that polysaccharides show high potential in achieving colon-specific drug delivery. [Abstract/Link to Full Text]

Takekuma Y, Kakiuchi H, Yamazaki K, Miyauchi S, Kikukawa T, Kamo N, Ganapathy V, Sugawara M
Difference between pharmacokinetics of mycophenolic acid (MPA) in rats and that in humans is caused by different affinities of MRP2 to a glucuronized form.
J Pharm Pharm Sci. 2007;10(1):71-85.
PURPOSE: Mycophenolic acid (MPA), an immunosuppressant, is excreted as its glucuronized form, MPAG. In humans, MPAG is mostly excreted into urine, whereas more than 80% of the dose is excreted into bile in rats. The aim of this study was to clarify the cause of the species difference. We investigated whether MPAG is a substrate of human organic anion transporters (hOATs), and we compared the affinities of multi-drug resistance-associated protein 2 (MRP2) for MPAG in rats and humans. METHODS: The inhibitory effects of MPAG on the uptake of typical substrates via hOAT1 and hOAT3 were determined using HeLa cells heterologously expressing hOAT1 and Xenopus laevis oocytes heterologously expressing hOAT3. MPAG transport activity via hOAT1 and hOAT3 was determined by the two-microelectrode voltage-clamp technique using Xenopus laevis oocytes expressing hOAT1 and hOAT3. The affinities of MPAG for hMRP2 and rMrp2 were determined by the inhibitory effects of MPAG on p-aminohippuric acid (a typical substrate) uptake using membrane vesicles expressing hMRP2 or rMrp2. RESULTS: MPAG inhibited the uptake of PAH via hOAT1 and hOAT3, and calculated IC50 values were 222.6+/-26.6 microM and 41.5+/-11.5 microM, respectively. However, MPAG was not transported by hOAT1 and hOAT3. MPAG strongly inhibited the uptake of PAH via both rMrp2 and hMRP2. However, the magnitudes of inhibitory effects were different. The calculated IC50 values were 286.2+/-157.3 microM and 1036.8+/-330.5 microM, respectively. CONCLUSION: MPAG is not a substrate but is an inhibitor of hOAT1 and hOAT3. The affinity of rMRP2 to MPAG was about 3.6 times as high as that of hMRP2. Therefore, the difference of affinity between hMRP2 and rMrp2 is a possible mechanism of the difference of excretion ratio of MPAG between rats and human. [Abstract/Link to Full Text]

Zhao Y, Gao J, Sun X, Chen H, Wu L, Liang W
Enhanced nuclear delivery and cytotoxic activity of hydroxycamptothecin using o/w emulsions.
J Pharm Pharm Sci. 2007;10(1):61-70.
PURPOSE: Hydroxycamptothecin-Emulsions (HCPT-E), being a promising delivery system for intravenous applications, were prepared. Here we evaluated the relation of cytotoxicity and intracellular drug amounts. The mechanism of the enhanced antiproliferative activity was also investigated. METHODS: In vitro cytotoxic potential were assessed by the methyl thiazol tetrazolium (MTT) assay. The capability of emulsions to preserve lactone form of HCPT and the time course of subcellular (cytoplasm and nuclei) drug distribution in vitro of HCPT-E were quantified by a reversed-phase HPLC assay. RESULTS: MTT results showed that the antiproliferative activity of HCPT-E was stronger than that of HCPT-Injections (HCPT-I) which is presently used in clinic. IC50 values of HCPT-E was much lower, by 4 and 7 times than those of HCPT-I. HeLa cells treated with HCPT-E for 4 h exhibited a considerable morphology change and displayed a hint of apoptotic cell death at 72 h. The lactone is the key position in HCTP molecule which is sensitive to pH. Lactone ring of HCPT in HCPT-E was stable for longer time than that of HCPT-I in the presence of human plasma or culture medium. Both the nuclear and cytoplasmic level of HCPT in HeLa cells exposed to HCPT-E at each time were higher by 18 to 33 times than those treated with HCPT-I. Drug amounts of nuclei was higher than that of cytoplasm in HeLa cells incubated with HCPT-E while it was reverse in HeLa cells exposed to HCPT-I. CONCLUSIONS: HCPT-E improved stability of HCPT, enhanced uptake, changed its intracellular distribution in favor of accumulating in the intracellular targets. [Abstract/Link to Full Text]

Wang YG, Song XJ, Feng SW, Ge YL, Yang JJ, He LL
Hyperthyroidism patients have shorter onset and duration time of rocuronium than euthyroidism patients.
J Pharm Pharm Sci. 2007;10(1):53-60.
PURPOSE: The possibility of altered response to nondepolarizing muscle relaxants in hyperthyroidism patients has not been documented. The present study was conducted to observe the onset and duration of rocuronium induced neuromuscular blockade in hyperthyroidism patients. METHODS: Eighteen hyperthyroidism patients undergoing subtotal thyroidectomy (H group) and eighteen euthyroidism patients undergoing operation on neck (E group) were studied. Anesthetized with propofol and fentanyl, all patients received rocuronium 0.6 mg/kg. The twitch high of adductor pollicis muscle was monitored by acceleromyography. The onset, the duration of the initial dose, the durations of the repeated maintenance doses, the repeated times, and the total doses of rocuronium were observed. RESULTS: The onset time of rocuronium in H group was significantly shorter than that in E group (P<0.05). The duration of the initial dose as well as the durations of the repeated maintenance doses in H group was significantly shorter than that in E group (P<0.05). The repeated times and the total dosage of rocuronium in H group were significantly more than that in E group (P<0.05). CONCLUSIONS: Hyperthyroidism patients experience a shorter onset time, a shorter duration, and require larger doses of rocuronium than euthyroidism patients. [Abstract/Link to Full Text]

Lee JH, Lee MG
Dose-dependent pharmacokinetics of telithromycin after intravenous and oral administration to rats: contribution of intestinal first-pass effect to low bioavailability.
J Pharm Pharm Sci. 2007;10(1):37-50.
PURPOSE: To evaluate the pharmacokinetics of telithromycin after intravenous and oral administration and to find the reason for incomplete F value (first pass-effect) after intravenous, intraportal, intragastric, and intraduodenal administration to rats. METHODS: Telithromycin was administered intravenously or orally at doses of 20, 50, and 100 mg/kg to rats. And hepatic, gastric, and intestinal first-pass effects of telithromycin were also measured after intravenous, intraportal, intragastric, and intraduodenal administration at a dose of 50 mg/kg to rats. RESULTS: The dose-normalized AUC values of telithromycin were dose-dependent (increased with increasing doses) after both intravenous and oral dose ranges studied, possibly due to saturable metabolism of telithromycin. After oral administration (50 mg/kg), approximately 4.06% of oral dose was not absorbed, F was approximately 27.5%, and the intestinal first-pass effect was approximately 63.4% of oral dose. The first-pass effects of telithromycin in the lung, heart, stomach, and liver were almost negligible, if any, in rats. CONCLUSIONS: The low F of telithromycin at a dose of 50 mg/kg was mainly due to considerable intestinal first-pass effect, approximately 63.4% of oral dose, in rats. [Abstract/Link to Full Text]

Arellano C, Philibert C, Vachoux C, Woodley J, Houin G
The metabolism of midazolam and comparison with other CYP enzyme substrates during intestinal absorption: in vitro studies with rat everted gut sacs.
J Pharm Pharm Sci. 2007;10(1):26-36.
PURPOSE: The purpose of this study was to quantify the intestinal metabolism of midazolam, a CYP P450 substrate, usually used as a probe for the activity of the isoform CYP3A4/1 and to compare it with previous results obtained for other P450 substrates such as testosterone, dextromethorphan and bupropion, which show some specificities for different CYP isoforms. The aim was to shed light on the role of metabolism in the intestinal tissues and the relationship with efflux mechanisms, such as by P-glycoprotein (P-gp) and the influence of metabolism on bioavailability. METHODS: We used the improved everted rat gut sac model to study in vitro the absorption and metabolism of the different CYP isoenzyme probes: midazolam, testosterone, bupropion and dextromethorphan. This method enables drug metabolism to be studied during absorption, conditions which mimic the in vivo situation. The drugs and their metabolites were measured by LC-MS in the mucosal and serosal media and in the mucosal tissue, to give a complete picture of the transport and metabolism. RESULTS: Midazolam, as with the other CYP probes, was metabolized in everted gut sacs. The metabolites were detected in the same proportions in both the serosal and mucosal compartments for midazolam, testosterone and bupropion. In the case of dextromethorphan, the metabolite methoxymorphinan was found at a higher concentration in the mucosal compartment, indicating efflux from the cells. The transport of dextromethorphan and its metabolite was not modified in the presence of verapamil, a P-gp inhibitor, thus demonstrating that dextromethorphan and methoxymorphinan were not P-gp substrates. CONCLUSION: Given that the rat is a widely used species for pre-clinical studies, the everted gut sac model provides a useful tool to assess the role of metabolism during drug absorption by the intestine and is also capable of demonstrating P-glycoprotein mediated transport. [Abstract/Link to Full Text]

Wang T, Gu X
In vitro percutaneous permeation of the repellent DEET and the sunscreen oxybenzone across human skin.
J Pharm Pharm Sci. 2007;10(1):17-25.
PURPOSE: DEET and oxybenzone are two essential active ingredients in repellent and sunscreen products. The percutaneous permeation of the two compounds across human skin from five commercially available repellent and sunscreen products was investigated in vitro. METHODS: Diffusion studies were carried out at 37 degrees C, using Franz-style diffusion cells and human epidermis (380 microm in thickness). The test products were evaluated either individually or in various combinations for up to 6 hours. Concentrations of both compounds permeated through the skin were measured using an HPLC assay. Permeability and permeation percentage of DEET and oxybenzone from different application approaches were calculated and statistically compared. RESULTS: The accumulated transdermal permeation was 0.5-25.7% for DEET and 0.3-1.6% for oxybenzone, respectively. Repellent lotion produced an 18-fold increase in transdermal permeation in comparison to that of repellent spray, while using repellent spray prior to sunscreen lotion resulted in the highest penetration of DEET among the study groups. Premixing sunscreen lotion with repellent spray at different ratios also produced significantly higher permeation of oxybenzone across the skin than the control, but other application approaches did not differentiate from the single sunscreen lotion. CONCLUSION: It was concluded from this study that human skin was less permeable to DEET and oxybenzone than artificial membranes, but was comparable to pig skin in permeability. DEET permeated transdermally more across human skin than oxybenzone, and both compounds acted as permeation enhancers when used simultaneously. Premixing repellent and sunscreen enhanced the overall penetration of both DEET and oxybenzone. Using different application sequences and amounts resulted in variable percutaneous permeation of DEET and oxybenzone through the skin. [Abstract/Link to Full Text]

Brennan B, Chiu Y, Berthelon L, Kolis S, Davies B
Effect of age and gender on the pharmacokinetics of R667, a novel agent for the treatment of emphysema, in healthy volunteers.
J Pharm Pharm Sci. 2007;10(1):9-16.
The purpose of the present study was to assess the impact of age and gender on the pharmacokinetics (PK) of R667. Thirty six healthy male and female volunteers (12 m 18-45 years; 12 m and 12 f > or = 65 years) received a single 1 mg oral dose of R667. Serial blood samples were collected for determination of plasma R667 and metabolite concentrations. The PK parameters of R667 were similar between elderly males and young males (Cmax = 9.8 vs 9.8 ng/mL; AUC(0-last) = 50.9 vs 47.3 ng x h/mL, respectively). Exposure of R667 increased in elderly females compared to elderly males (Cmax = 13.1 vs 9.8 ng/mL; AUC(0-last) = 60.8 vs 47.3 ng x h/mL, respectively). When the CL/F was corrected for BSA and V/F corrected for weight these differences were no longer evident. In conclusion these exposure differences are not considered clinically relevant, and no dose adjustment of R667 is required based on gender or age. [Abstract/Link to Full Text]

Kim TY, Jung DI, Kim YI, Yang JH, Shin SC
Anesthetic effects of lidocaine hydrochloride gel using low frequency ultrasound of 0.5 MHz.
J Pharm Pharm Sci. 2007;10(1):1-8.
PURPOSE: Low-frequency ultrasound has a significant effect on the transdermal permeation of high molecular weight drugs. However, the rate of permeation in pulsed mode is quite low necessitating considerable time to apply the ultrasound. 0.5 MHz ultrasound, which is a relatively higher frequency in the low-frequency range, can be applied in high intensity in continuous mode. METHODS: A transducer was used to administer an anesthetic drug transdermally on healthy volunteers. The anesthetic effect was measured following administration on placebo, lidocaine HCl alone and lidocaine HCl with 0.5 and 1.0 MHz ultrasound (n=8/group). RESULTS: In surface anesthesia, the phonophoresis group showed a significantly higher pain threshold than the other groups but there was no significant difference between the phonophoresis groups according to the ultrasound frequency. In conduction anesthesia, the 0.5 MHz phonophoresis group showed a significant change in their pain threshold and amplitude of sensory nerve action potential (SNAP) compared with the other groups. CONCLUSIONS: Although there are limitations in applying 0.5 MHz ultrasound in phonophoresis for conduction anesthesia using lidocaine hydrochloride for a nerve block, it is more effective than the 1 Mhz that is widely used in clinical situations.Carbamazepine is a poor water soluble drug and its bioavailability is limited by dissolution rate. Dissolution, serum concentration and anticonvulsive effect of the drug have been evaluated after cogrinding with microcrystalline cellulose. A cogrinding technique was used to increase the dissolution, serum concentrations and anticonvulsive effect of the drug. A novel deconvolution technique of in vitro-in vivo correlation was evaluated. [Abstract/Link to Full Text]

Cruciol-Souza JM, Thomson JC
Prevalence of potential drug-drug interactions and its associated factors in a Brazilian teaching hospital.
J Pharm Pharm Sci. 2006;9(3):427-33.
PURPOSE: The hazards of prescribing many drugs, including side-effects, drug-drug interactions (DDI) and difficulties of compliance have long been recognized as particular problems when prescribing. This study estimates the rate and factors associated with potential DDI in prescriptions from wards of a Brazilian teaching hospital. METHODS: Data were retrieved from wards of a teaching hospital (300 beds) handwritten prescription, once a week during a period of 4 months in 2004. Potential DDI were identified using DrugReax system. Patient's age and gender, number of prescribers; number of drugs and therapeutic drug classes on prescriptions were explored as associated factors to DDI. RESULTS: The overall frequency of potential DDI was 49.7%. The frequency of the potentially major DDI was 3.4%, with digoxin-hydrochlorothiazide as the most common interacting pair. The rate of potential DDI was significantly associated to in-patients' gender [woman, Odds ratio (OR)=1.23 (P=0.035)], age=55 years old [OR=1.5 (P=0.0008)], number of therapeutic drug class (ATC code, level 1)=4 [OR=5.5 (P=0.0000), cardiology patients [OR=7.87 (P=0.0000)] hospitalized at weekends [OR=1.24 (P=0.039)] and having digoxin prescribed [OR=16.79 (P=0.0000)]. A positive correlation was found between DDI, patient's age, number of drugs and therapeutic action ATC codes were significant, controlling for gender (Pearson's r=0.628, P=0.001). CONCLUSIONS: Cardiology women inpatients, age more then 55 years old, 7 or more drugs prescribed (including digoxin) and hospitalized at weekends should be closely monitored for adverse outcomes from DDI. A collaborative approach toward drug selection is strongly recommended, as well as electronic prescribing and development of pharmaceutical care in Brazilian hospitals. [Abstract/Link to Full Text]

Glass BD, Haywood A
Stability considerations in liquid dosage forms extemporaneously prepared from commercially available products.
J Pharm Pharm Sci. 2006;9(3):398-426.
The pharmacist, both in community and hospital pharmacy practice, is often challenged with the preparation of a liquid dosage form not available commercially for paediatric patients, those adults unable to swallow tablets or capsules and patients who must receive medications via nasogastric or gastrostomy tubes. Recognising the lack of information available to healthcare professionals, a general discussion of the various parameters that may be modified in preparing these dosage forms and a tabulated summary of the dosage forms presented in the literature is described, which, although not exhaustive, will provide information on the formulation and stability of the most commonly prepared extemporaneous liquid dosage forms. An extensive survey of the literature and investigation of 83 liquid dosage forms revealed that stability considerations were of concern for only 7.2% of these liquid dosage forms, extemporaneously prepared from the following commercially available products: captopril, hydralazine hydrochloride, isoniazid, levothyroxine sodium, phenoxybenzamine hydrochloride and tetracycline hydrochloride. Inclusion of the antioxidant, sodium ascorbate in the liquid dosage form for captopril resulted in improved stability at 4 degrees C. Hydralazine hydrochloride, isoniazid and phenoxybenzamine hydrochloride were adversely affected due to interactions with excipients in the formulation, while the effect of the preservative in lowering the pH in a levothyroxine sodium mixture resulted in decreased stability. Interestingly, the instability in these formulations is primarily due to interactions between the drug substance and the excipients rather than degradation of the active pharmaceutical ingredient by standard routes such as oxidation, hydrolysis, photolysis or thermolysis. This low percentage however illustrates the low risk associated with these dosage forms investigated. It may be concluded that when considering the safety and efficacy of liquid dosage forms prepared extemporaneously, it is thus important to consider not only the stability of the drug substance but the entire formulation. [Abstract/Link to Full Text]

Pospisilova N, Semecky V, Jamborova G, Pospechova K, Solichova D, Andrys C, Zdansky P, Nachtigal P
Endoglin expression in hyper-cholesterolemia and after atorvastatin treatment in apoE-deficient mice.
J Pharm Pharm Sci. 2006;9(3):388-97.
PURPOSE: Endoglin (CD105) is a marker of activated endothelium and a modulator of TGF-beta signaling. We hypothesized whether endothelial expression of endoglin is changed in hypercholesterolemia as well as whether its expression is affected by atorvastatin treatment in apoE-deficient mice. METHODS: ApoE-deficient mice were fed with the chow diet for either 4 weeks or for 12 weeks respectively. In two treated groups, mice were fed with chow diet except atorvastatin was added to the diet for the last 4 weeks or for the last 8 weeks respectively, before euthanasia. RESULTS: Administration of atorvastatin did not affect lipid parameters after 4 weeks treatment, however increased all lipid parameters after 8 weeks of treatment. Stereological analysis of immunohistochemical staining revealed that atorvastatin significantly decreased endoglin expression in endothelium after 4 weeks of treatment but increased it after 8 weeks of treatment. CONCLUSIONS: This study demonstrates that endoglin is expressed by aortic endothelium showing similar staining patterns like other markers involved in the process of atherosclerosis. In addition, we showed that endoglin expression in endothelium could be affected by the administration of atorvastatin beyond its lipid lowering effects in apoE-deficient mice. [Abstract/Link to Full Text]

Mobach MP
The merits of a robot: a Dutch experience.
J Pharm Pharm Sci. 2006;9(3):376-87.
PURPOSE: To determine the merits of a robot at the community pharmacy in a quasi-experiment. METHOD: The applied methods for data-collection were barcode-time measurements, direct observations, time-interval studies, and tally at a Dutch community pharmacy. The topics consisted of workload, waiting times, congestion, slack, general work, counter work, and work at the consultation room. The topics were studied in pre-test and post-test stages, each stage during six weeks. By using these topics and some additional data from the pharmacy, the economics of the robot were also assessed. RESULTS: The workload decreased with 15 prescriptions per person per day. The waiting times decreased with one minute and 18 seconds per dispensing process, reducing the wait until counter contact. The day congestion decreased with one hour 27 minutes and 36 seconds, and the day slack increased with 28 minutes. The analysis of the general work showed no appreciable difference in the bulk of the care-related activities and the other activities. However, some work was re-shuffled: 7% increase at counter work and 7% decrease at logistics. Moreover, statistically significant increases were observed at counter work (5%) and robot work (4%), and significant decreases at telephone (3%) and filling work in presence of the patient (4%). The counter tally study showed a rise in care-related activities with 8%. Moreover, it also illuminated a statistically significant decrease at no information (11%) and an increase at only social (2%). The consultation room was never used during the study. The pharmacy economics of the robot showed that the robot had high estimated costs for purchase, depreciation, and maintenance: EUR 187,024 in the first year. Moreover, the robot had positive impact on waiting times, congestion, staffing, logistics, and care-related work, which was estimated on EUR 91,198 in the first year. The estimated payback time of the robot was three years. CONCLUSIONS: An introduction of the robot may indeed have the often supposed positive effects on pharmaceutical care. Even though the costs are high and the technical problems are present, the robot seems to be financial beneficial after three years. The robot can create space for pharmaceutical care, but it has a substantial cost. [Abstract/Link to Full Text]

Ye P, Sheng L, Zhang C, Liu Y
Atorvastatin attenuating down-regulation of peroxisome proliferator-activated receptor gamma in preventing cardiac hypertrophy of rats in vitro and in vivo.
J Pharm Pharm Sci. 2006;9(3):365-75.
PURPOSE: To investigate whether the role of atorvastatin in suppression of cardiac hypertrophy is potentially associated with the change of peroxisome proliferator-activated receptor gamma (PPARgamma) expression, and the anti-inflammatory effect in vitro and in vivo. METHOD: Cardiac hypertrophy was established by angiotensin II in neonatal cardiac myocytes in vitro and incomplete ligation of abdominal aorta of SD rats in vivo. PPARgamma and cytokines mRNA expression was evaluated by RT-PCR, and the rate of protein synthesis in cardiac myocytes by 3H-leucine incorporation. RESULTS: Atorvastatin attenuated downregulation of PPARgamma mRNA and inhibited brain natriuretic peptide (BNP), interleukin-1beta (IL-1beta) and matrix metalloproteinase 9 (MMP9) mRNA expression, as well as 3H-leucine incorporation in a dose-dependent manner in vitro. Furthermore, atorvastatin reduced the mRNA expression of BNP, IL-1beta and MMP9, and enhanced PPARgamma mRNA expression, and diminished the pressure overload-induced increase in the ratio of heart weight to body weight, left ventricular wall thickness and myocyte diameter of rats in vivo. CONCLUSION: Atorvastatin prevents cardiac hypertrophy of rats, probably associated with the modulation of PPARgamma and the inhibition of myocardial inflammation. Atorvastatin may play a role in prevention and treatment of cardiovascular diseases characterized by cardiac hypertrophy. [Abstract/Link to Full Text]

Aghazadeh-Habashi A, Ibrahim A, Carran J, Anastassiades T, Jamali F
Single dose pharmacokinetics and bioavailability of butyryl glucosamine in the rat.
J Pharm Pharm Sci. 2006;9(3):359-64.
PURPOSE: To study single dose pharmacokinetics and bioavailability of the synthetic glucosamine analogue, N-butyryl glucosamine (GLcNBu) after different routes of administration, and also the effect of food following oral doses of GLcNBu in the rat, and stability and absorption of GLcNBu across the rat everted intestine. METHOD: GLcNBu was administered intravenously (i.v.), intraperitoneally (i.p.) and orally. Effect of food was studied following oral administration only. Single doses of 223 mg/kg were administered in all cases. Serial blood samples were collected from the jugular vein for GLcNBu determination. Everted excised rat intestine segments were suspended in Tris buffer at 37 degrees C and samples collected from both serosal and mucosal sides. GLcNBu was measured using HPLC. RESULT: Following i.v. administration, the terminal half-life was 0.29 -/+ 0.06 h, volume of distribution at steady state was 2.1 -/+ 0.26 L/kg and total body clearance was 5.23 -/+ 1.44 L/h/kg. Bioavailability was less than 17% and 100% following oral and i.p. doses respectively. GLcNBu was rapidly absorbed after oral doses (Tmax, 29-40 min). Food had no significant effect on the pharmacokinetics of GLcNBu. There was no evidence of breakdown of GLcNBu in the presence of everted intestine. The mucosal to serosal transport of GLcNBu was about 20% after 2 h incubation. CONCLUSION: GLcNBu has rapid but low absorption and is widely distributed and efficiently cleared. The gut rather than liver is mainly responsible for the low bioavailability of GLcNBu. Limited absorption of GLcNBu suggests a transport dependent absorption. Food does not significantly affect the bioavailability of GLcNBu. [Abstract/Link to Full Text]

Escobar-Chávez JJ, López-Cervantes M, Naďk A, Kalia YN, Quintanar-Guerrero D, Ganem-Quintanar A
Applications of thermo-reversible pluronic F-127 gels in pharmaceutical formulations.
J Pharm Pharm Sci. 2006;9(3):339-58.
It is, sometimes, desirable to maintain a constant plasma drug concentration within the therapeutically effective concentration range. The use of high viscosity hydromiscible vehicles such as hydrophilic gels, is one of various approaches for controlled drug delivery, and represents an important area of pharmaceutical research and development. Of these systems, Pluronic F-127 (PF-127) provides the pharmacist with an excellent drug delivery system for a number of routes of administration and is compatible with many different substances. Gels containing penetration enhancers have proven to be especially popular for administering anti-inflammatory medications since they are relatively easy to prepare and very efficacious. [Abstract/Link to Full Text]

Asghar LF, Chandran S
Multiparticulate formulation approach to colon specific drug delivery: current perspectives.
J Pharm Pharm Sci. 2006;9(3):327-38.
Colon specific drug delivery has gained increased importance not just for the delivery of drugs for the treatment of local diseases associated with the colon but also as potential site for the systemic delivery of therapeutic peptide and proteins. To achieve successful colon targeted drug delivery, a drug needs to be protected from degradation, release and/or absorption in the upper portion of the GI tract and then ensure abrupt or controlled release in the proximal colon. Drug modifications through covalent linkages with carrier or prodrug approach and formulation based approaches can be used for colonic delivery. Report suggests that drug carrier systems larger than 200 mm possess very low gastric transit time due to physiological condition of the bowel in colitis. And for this reason and considering the selective uptake of micron or sub-micron particles by cancerous and inflamed cells/ tissues a multiparticulate approach based on pellets, granules, microsphere or nanoparticle type formulation is expected to have better pharmacological effect in the colon. The review is aimed at understanding recent advancements made in multiparticulate formulation approach for colon specific delivery of medicaments. [Abstract/Link to Full Text]

Peterson ML, Hickey MB, Zaworotko MJ, Almarsson O
Expanding the scope of crystal form evaluation in pharmaceutical science.
J Pharm Pharm Sci. 2006;9(3):317-26.
The commentary seeks to provide a brief history and perspective on the importance of crystal forms of pharmaceuticals as a means of achieving performance criteria. The expanding scope of crystal form selection, emergence of crystal engineering in pharmaceutical science and pharmaceutical co-crystals are topics of this brief review. [Abstract/Link to Full Text]

Barzegar-Jalali M, Nayebi AM, Valizadeh H, Hanaee J, Barzegar-Jalali A, Adibkia K, Anoush M, Sistanizad M
Evaluation of in vitro-in vivo correlation and anticonvulsive effect of carbamazepine after cogrinding with microcrystalline cellulose.
J Pharm Pharm Sci. 2006;9(3):307-16.
PURPOSE: Carbamazepine is a poor water soluble drug and its bioavailability is limited by dissolution rate. Dissolution, serum concentration and anticonvulsive effect of the drug have been evaluated after cogrinding with microcrystalline cellulose. A cogrinding technique was used to increase the dissolution, serum concentrations and anticonvulsive effect of the drug. A novel deconvolution technique of in vitro in vivo correlation was evaluated. METHODS: The drug coground with microcrystalline cellulose, the corresponding physical mixture, unground and ground drug powder were subjected to dissolution measurement. Coground and unground drug serum concentrations were investigated in rabbits. Also the anticonvulsive effects of the latter preparations were assessed in mice. For elucidation of observed in vitro and in vivo differences FT-IR spectroscopy, X-ray diffraction patterns and DSC thermograms of the preparations were studied. RESULTS: The dissolution of the coground was the highest (percent dissolved in the first 20 minutes, %D20', was 97.5). The unground drug powder exhibited the lowest dissolution (%D20'=40). The difference was reflected in their corresponding area under the mean serum concentration curves between 0-16 hr (118.96 vs 54.17 microg x hr/ml) as well as protection abilities against strychnine and electrically induced seizures. The onset of tonic seizures induced by strychnine was increased between 40-140% in the case of the coground system depending on dose and time of carbamazepine administration. CONCLUSION: Cogrinding was an effective technique in increasing carbamazepine dissolution due to reduced crystallinity as seen in X-ray pattern, enhanced wettability and decreased particle size, which in turn resulted in increased serum concentrations and its anticonvulsive effect. A novel simple deconvolusion technique not requiring intravenous data denoted as the double reciprocal area method was used to establish correlation between in vitro and in vivo parameters. [Abstract/Link to Full Text]

Kulmatycki KM, Jamali F
Drug disease interactions: role of inflammatory mediators in depression and variability in antidepressant drug response.
J Pharm Pharm Sci. 2006;9(3):292-306.
Increased cytokine expression and concurrent psychiatric symptoms were initially observed after administration of cytokines to patients afflicted with cancer, hepatitis and multiple sclerosis. Cytokines are a diverse group of soluble messenger proteins involved in the regulation, repair of cells, and control of immune events. During an inflammatory event expression of CD4+ T-lymphocyte helper (Th)1 cells that primarily produce pro-inflammatory cytokines is favored which can lead to development of inflammatory disease (e.g., cardiovascular disease). Similarly, relationships have been shown to exist between changes in inflammatory mediator concentrations, specifically pro-inflammatory cytokines, and depression. An increased prevalence of depression in patients afflicted with co-morbid inflammatory disease indirectly supports this association. In further support, antidepressants have been suggested to alleviate symptoms of depression via anti-inflammatory actions. Administration of anti-cytokines to patients with concurrent depression and inflammatory disease has resulted in relief of depressive symptoms. The exact role of inflammation in development of depression, however, remains to be determined. Nevertheless, increased expression of inflammatory mediators in depressed patients occurs which may lead to variability in response to antidepressant drug therapy. For example, depressed patients non-responsive to drug treatment are reported to have increased cell mediated immunity shown by elevated CD4+ T-cell activity, pro-inflammatory cytokine expression, and stimulation of the acute phase response. This suggests a psycho-neuroimmunological approach may be required for optimal pharmacotherapy. [Abstract/Link to Full Text]

Megalli S, Davies NM, Roufogalis BD
Anti-hyperlipidemic and hypoglycemic effects of Gynostemma pentaphyllum in the Zucker fatty rat.
J Pharm Pharm Sci. 2006;9(3):281-91.
Gynostemma pentaphyllum is a traditional Chinese medicine used for a variety of conditions, including elevated cholesterol. We have examined the pharmacological anti-hyperlipidemic and hypoglycemic effectiveness of Gynostemma pentaphyllum in the obese Zucker fatty diabetic rat model. After treatment for 4 days Gynostemma pentaphyllum 250 mg/kg reduced triglyceride (33%), total cholesterol, (13%) and low density lipoprotein cholesterol levels (33%). These effects were dose-dependent and maintained for at least 5 weeks. Chronic treatment for 3-5 weeks also reduced post-prandial hypertriglyceridemia induced by olive oil 10 mg/kg in the Zucker fatty rats but had no significant effect in lowering sucrose-induced hyperglycemia in Sprague-Dawley rats. A novel regulation by Gynostemma of glucose levels was also observed in the Zucker fatty rat model. In a glucose tolerance test in obese and lean Zucker rats pretreatment with Gynostemma pentaphyllum 250 mg/kg demonstrated glucose levels were significantly less 2 hours post challenge (20%) in the Gynostemma pentaphyllum obese rats compared to the control group. Gynostemma pentaphyllum did not significantly reduce glucose levels at 120 min in the lean strain, in contrast to the 20% decrease seen in the obese rat. In vitro, Gynostemma pentaphyllum inhibited alpha-glucosidase activity (50% inhibition at 42.8), which compared to acarbose (50% at 53.9 microg/mL). The improvement in glucose tolerance at 120 min by Gynostemma pentaphyllum in obese Zucker fatty rats but not lean rats suggests that it may improve insulin receptor sensitivity and together with the significant reduction of hypertriglyceridemia, cholesterol and low density lipoprotein cholesterol suggests that Gynostemma should be examined further by oral hypoglycemic/anti-hyperlipidemic therapy. [Abstract/Link to Full Text]

Trubetskoy OV, Finel M, Burke TJ, Trubetskoy VS
Evaluation of synthetic polymeric micelles as a stabilization medium for the handling of membrane proteins in pharmaceutical drug discovery.
J Pharm Pharm Sci. 2006;9(3):271-80.
PURPOSE: Polymeric micelles have been used for solubilization of insoluble drugs and as carriers for drug delivery applications. Here we evaluated an application of the synthetic polymeric micelles in experiments designed to improve the handling and stability of membrane proteins targets. METHODS: Particle sizing by dynamic light scattering was performed in a Zeta Plus Photon Correlation Spectrometer at 532 nm. UGT1A1 activity has been measured in fluorescent assay using scopoletin as a substrate. COX-2 activity has been measured in a fluorescent assay using Amplex Red. Fluorescence Resonance Energy Transfer (FRET) was monitored using either 463 nm excitation wavelength (the emission range 500-600 nm) or 395 nm excitation wavelength (the emission range 500-600 nm). RESULTS: Incorporation of membrane proteins into PreserveX-QML polymeric micelles resulted in improved homogeneity and stability of the preparation and in reduced light scattering. Stabilization of the biological activity of micelle-incorporated membrane proteins, such as the human UGT1A1 and COX-2 both during extended incubations at room temperature and during multiple freeze/thaw cycles, has been achieved. CONCLUSION: PreserveX-QML polymeric micelles help to homogenize and disperse membrane proteins preparations and stabilize the biological activity of the proteins making it more suitable for pharmaceutical assays and applications. [Abstract/Link to Full Text]

Jouyban A, Acree WE
In silico prediction of drug solubility in water-ethanol mixtures using Jouyban-Acree model.
J Pharm Pharm Sci. 2006;9(2):262-9.
PURPOSE: A predictive method was proposed to predict solubility of drugs in water-ethanol mixtures at various temperatures based on the Jouyban-Acree model. The model requires the experimental solubility data of the drug in mono-solvent systems. METHODS: The accuracy of the proposed prediction method was evaluated using collected experimental solubility data from the literature. The proposed method is: log Xm,T = fc log Xc,T + fw log Xw,T + fcfw[724.21/T + 485.17(fc-fw)/T + 194.41(fc-fw)2/T] Where Xm,T, Xc,T and Xw,T are the solute solubility at temperature (T) in mixed solvent and neat cosolvent and water, respectively, fc and fw denote the solute free fraction of cosolvent (ethanol) and water. The average absolute error (AAE) of the experimental and the predicted solubilities was computed as an accuracy criterion and compared with that of a well-established log-linear model. RESULTS: The AAE (+/-SD) of the Jouyban-Acree and log-linear models were 0.19 (+/-0.13) and 0.48 (+/-0.28), respectively. The mean difference of AAEs was statistically significant (p < 0.0005) revealing that the Jouyban-Acree model was provided more accurate predictions. Although the log-linear model was used to predict solubility at a fixed temperature (25 or 23 degrees C), the results also showed that the model could be employed to predict the solubility in solvent mixtures at various temperatures. CONCLUSION: More accurate predictions were provided using the Jouyban-Acree model in comparison with a previously established log-linear model of Yalkowsky. The prediction methods were successfully extended to predict the solubility in water-ethanol mixtures at various temperatures. [Abstract/Link to Full Text]

Yeung PK, Feng JD, Fice D
Exercise hemodynamic and neurohormone responses as sensitive biomarkers for diltiazem in rats.
J Pharm Pharm Sci. 2006;9(2):245-51.
PURPOSE: To investigate the potential of exercise hemodyanamic and neurohormone variables as sensitive biomarkers for pre-clinical evaluation of diltiazem (DTZ). METHODS: Sprague Dawley (SD) rats were randomly divided into 3 groups (n = 6 - 8 each), and each group received DTZ 10 mg/kg twice daily for 5 doses or saline followed by a treadmill exercise protocol for 7 min with speed set at 7 m/min at 3 % grade. The 3rd group received saline but no exercise. RESULTS: Exercise increased SBP from 108 +/- 2 to 131 +/- 3 mmHg, and HR from 437 +/- 6 to 503 +/- 6 bpm, and plasma epinephrine concentrations from 2.0 +/- 0.6 to 5.8 +/- 1.7 ng/mL in control rats (p < 0.05 for all variables), but had no significant effect on DBP (81 +/- 5 vs 87 +/- 6 mmHg) and plasma norepinephrine concentrations (1.5 +/- 0.2 vs 3.9 +/- 0.4 ng/mL). The hemodynamic responses to exercise were significantly attenuated by DTZ (p < 0.05), but the effect on neurohormone response was minimal (p > 0.05). CONCLUSION: Exercise hemodynamic and neurohormone responses are sensitive biomarkers which could be used for safety and efficacy evaluation of DTZ and perhaps also other calcium antagonists in pre-clinical animal models. [Abstract/Link to Full Text]

Li S, Shen Y, Li W, Hao X
A common profile for polymer-based controlled releases and its logical interpretation to general release process.
J Pharm Pharm Sci. 2006;9(2):238-44.
PURPOSE: The release of drug from a polymeric matrix is complicated. It often involves drug diffusion, interface movement and various interactions. It also shows a dependence on the length of polymer rod. With Lc as the 'critical length', three different phases can be distinguished from the kinetic process. Prior to reaching Lc, the interface movement plays a key role on determining the release. Otherwise, the drug diffusion can dominate the release process. Near Lc, however, both factors are involved. The rate of interface movement is closely associated with the time and position in the polymer rod. Taking these characters into account, a common model is presented in this article to be tentatively used to interpret the release process. These results provide preliminarily an insight into the understanding of controlled release process. [Abstract/Link to Full Text]

Marcel Musteata F, Pawliszyn J
Determination of free concentration of Paclitaxel in liposome formulation.
J Pharm Pharm Sci. 2006;9(2):231-7.
PURPOSE: An important step in the development of liposome-based formulations is estimating the free drug concentration in the aqueous solution surrounding liposomes. This research presents a new method for determination of free concentrations, based on membrane-protected solid-phase microextraction (SPME). METHODS: For effective direct extraction of low molecular weight compounds from complex liquid samples, a hollow membrane was used to form a concentric sheath around a coated SPME fiber. The membrane blocked the access of large particles, like liposomes, to the coating surface, while target analytes with low molecular weight diffused through the membrane and reached the extraction phase. Quantification was conveniently performed by reversed-phase liquid chromatography coupled to electrospray ionization mass spectrometry. RESULTS: The carbowax/templated resin SPME fiber was determined to be the most suitable for these assays, providing enough sensitivity when an extraction time of one hour was used. The free concentration of paclitaxel was found to be 0.36 microg/mL, significantly below the solubility limit of paclitaxel in water. CONCLUSION: The method was successfully applied for determining free paclitaxel in liposome formulations based on dioleyl-trimethyl-ammonium-propane, with good linearity over the range of concentrations of interest. The method was faster and more practical than equilibrium dialysis, as the SPME approach provided preconcentration and convenient delivery to the analytical system. [Abstract/Link to Full Text]

Recent Articles in Journal of Drug Targeting

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Recent Articles in Drug Metabolism and Pharmacokinetics

Ozawa S
Collaborative causes of individual variation in drug metabolism and pharmacokinetics and drug response: variations discovered from genetics and genomics approaches.
Drug Metab Pharmacokinet. 2007 Jun;22(3):141. [Abstract/Link to Full Text]

Hanzawa Y, Sasaki T, Hiratsuka M, Ishikawa M, Mizugaki M
Three novel single nucleotide polymorphisms (SNPs) of CYP2S1 gene in Japanese individuals.
Drug Metab Pharmacokinet. 2007 Apr;22(2):136-40.
We analyzed all nine exons and exon-intron junctions of the CYP2S1 gene in 200 Japanese individuals and identified the following three novel single nucleotide polymorphisms (SNPs): 4612G>A (Glu147Glu) in exon 3, 5478C>T (Leu230Leu) and 5479T>G (Leu230Arg, CYP2S1*5A) in exon 5. The allele frequencies were 0.013 for 4612G>A, 0.058 for 5478C>T, and 0.003 for 5479T>G. In addition, a known SNP 1324C>G (Pro74Pro) was detected at a frequency of 0.300. [Abstract/Link to Full Text]

Fukushima-Uesaka H, Saito Y, Maekawa K, Hasegawa R, Suzuki K, Yanagawa T, Kajio H, Kuzuya N, Noda M, Yasuda K, Tohkin M, Sawada J
Genetic variations of the ABC transporter gene ABCC3 in a Japanese population.
Drug Metab Pharmacokinet. 2007 Apr;22(2):129-35.
An ATP-binding cassette transporter, multidrug resistance-related protein 3 (MRP3), is encoded by the ABCC3 gene. The MRP3 protein is expressed in several tissues, and functions as an efflux transporter for conjugated as well as unconjugated substrates. In this study, the 31 ABCC3 exons and their flanking introns were comprehensively screened for genetic variations in 89 Japanese subjects. Forty-six genetic variations, including 21 novel ones, were found: 8 were located in the 5'-flanking region, 14 in the coding exons (8 synonymous and 6 nonsynonymous variations), and 24 in the introns. Of these 46 variations, five novel nonsynonymous variations, 2221C>T (Gln741Stop), 2395G>A (Val799Met), 2798_2799delAG (Gln933ArgfsX64), 3657C>A (Ser1219Arg), and 4217C>T (Thr1406Met), were found as heterozygous variations. The allele frequencies were 0.011 for Ser1219Arg and 0.006 for the other four variations. Gln741Stop induces a stop codon at codon 741. Gln933ArgfsX64 causes a frame-shift at codon 933, resulting in early termination at codon 997. Both variations result in loss of 6 transmembrane helices (from the 12th to 17th helices) in the C-terminus and all regions of nucleotide binding domain 2. Thus, both variant proteins are assumed to be inactive. These data provide fundamental and useful information for pharmacogenetic studies on MRP3-transported drugs in Japanese. [Abstract/Link to Full Text]

Tougou K, Fukuda T, Ito T, Yamazaki H, Fujio Y, Azuma J
Genetic polymorphism of bile acid CoA: amino acid N-acyltransferase in Japanese individuals.
Drug Metab Pharmacokinet. 2007 Apr;22(2):125-8.
In the present study, we identified three novel single nucleotide polymorphisms (SNPs), 147C>T in exon 2 (silent), 602G>C in exon 3 (Arg201Pro), and 1134C>T in exon 4 (silent), in the gene of bile acid CoA: amino acid N-acyltransferase (BAAT) by resequencing the entire coding region and the exon-intron junctions of 100 Japanese individuals. The allelic frequencies were 0.005 for 147C>T, 0.095 for 602G>C, and 0.015 for 1134C>T. The two known SNPs, 59G>A (Arg20Gln, rs1572983) and UTR1513G>A (rs2229594), were detected at a frequency of 0.500 and 0.425, respectively. In the haplotype analysis for the 59G>A and 602G>C polymorphisms, the allelic frequency of 59G-602G, 59G-602C, 59A-602G and 59A-602C was 0.405, 0.095, 0.500 and 0.000, respectively. On the other hand, the allelic frequency of the nonsynonymous SNP 602G>C was 0.194 in a Caucasian population. [Abstract/Link to Full Text]

Tayama Y, Moriyasu A, Sugihara K, Ohta S, Kitamura S
Developmental changes of aldehyde oxidase in postnatal rat liver.
Drug Metab Pharmacokinet. 2007 Apr;22(2):119-24.
In this study, the developmental changes and variability of aldehyde oxidase in postnatal rat liver were examined. Postnatal day 1, 7 and 14 rats showed little or no liver aldehyde oxidase activity, as evaluated in terms of the activities for oxidation of benzaldehyde to benzoic acid, N-1-methylnicotinamide (NMN) to N-1-methyl-2-pyridone-5-carboxamide (2-PY) and N-1-methyl-4-pyridone-3-carboxamide (4-PY), and methotrexate (MTX) to 7-hydroxymethotrexate (7-OH-MTX). However, these oxidase activities were markedly increased in liver cytosol from the rats after postnatal day 14. The activity was then maintained up to 6 weeks. The amounts of 2-PY and 4-PY formed from NMN were almost the same. The development of aldehyde oxidase activity toward benzaldehyde was closely correlated with that of oxidase activity toward NMN and MTX. The expression of aldehyde oxidase at postnatal day 14 was confirmed by Western blotting analysis. The density of bands of aldehyde oxidase was closely correlated with the oxidase activity toward benzaldehyde. The developmental changes of aldehyde oxidase activities during postnatal reflected the changes in the amount of the oxidase protein. Thus, aldehyde oxidase activity in rats rapidly increases from birth, reaching a plateau within 4 weeks, and is regulated by expression of the protein. [Abstract/Link to Full Text]

Nakamura K, Watanabe A, Okudaira N, Okazaki O, Sudo K
Effect of ion suppression on judgment of enzyme inhibition and avoidance of error by utilizing a stable isotope-labeled probe substrate: example of CYP3A4 inhibition with [13C4,15N] labeled midazolam as a substrate.
Drug Metab Pharmacokinet. 2007 Apr;22(2):113-8.
An advantage of using LC-MS(/MS) for in vitro CYP inhibition screening is that it does not require extensive sample preparation and chromatographic separation. Attention must be paid, however, to ion suppression effects on analytes caused by the test compound as well as endogenous compounds. In this study, we have shown the ion suppression of 1'-hydroxymidazolam (analyte) and dextrorphan (IS) by erythromycin, as an example, which may cause over- or underestimation of CYP3A4 inhibition. To avoid this kind of effect, we proposed to use a stable isotope-labeled substrate and determine labeled metabolites by using unlabeled authentic compounds of each metabolite. We showed that CYP3A4 activity was determined with high accuracy and precision by using stable isotope-labeled midazolam even in the presence of an ion suppressor at high concentrations in the samples. This method is useful not only for the CYP inhibition screening but also for testing drug candidates to predict changes in metabolite formation by the possible co-administered drugs. [Abstract/Link to Full Text]

Hara Y, Nakajima M, Miyamoto K, Yokoi T
Morphine glucuronosyltransferase activity in human liver microsomes is inhibited by a variety of drugs that are co-administered with morphine.
Drug Metab Pharmacokinet. 2007 Apr;22(2):103-12.
Morphine is an analgesic drug used for the treatment of acute and chronic pain syndromes for cancer patients. Glucuronidation is a major pathway of the elimination of morphine in humans. Morphine is metabolized to 3-glucuronide (no analgesic effect) and 6-glucuronide (more potently analgesic than morphine) mainly by UGT2B7. In the present study, we investigated the inhibitory effects of a variety of drugs on the morphine glucuronosyltransferase activities in human liver microsomes. Twenty-one drugs including anticancer drugs, immunosuppressants, analgesics, anticonvulsants, antidepressants, antipsychotic drugs were selected in this study, because they are frequently co-administered with morphine. We found that 10 out of 21 drugs, tamoxifen, tacrolimus, diclofenac, carbamazepine, imipramine, clomipramine, amitriptyline, diazepam, lorazepam and oxazepam extensively inhibited the morphine 3- and 6-glucuronosyltransferase activities. Although some of the drugs are not substrates of UGT2B7, they would be potent inhibitors of UGT2B7. If patients receive morphine and these drugs simultaneously, the drug-drug interaction may change the levels of morphine and these glucuronides, resulting in altered analgesic efficacy and the risk of side effects. The results presented here will assist clinicians in choosing the proper drugs and/or dosages, and enable them to anticipate potential drug-drug interactions. [Abstract/Link to Full Text]

Kato R, Yuasa H, Inoue K, Iwao T, Tanaka K, Ooi K, Hayashi Y
Effect of Lactobacillus casei on the absorption of nifedipine from rat small intestine.
Drug Metab Pharmacokinet. 2007 Apr;22(2):96-102.
Lactobacillus casei Shirota strain (L. casei) has a modulating effect on the production of cytokines, which often play important roles in drug metabolism, in the inflamed intestinal mucosa. We evaluated the effect of L. casei administered orally in advance for 4 weeks on the absorption of nifedipine from the rat small intestine. The maximum concentration of nifedipine in plasma after administration into the intestinal loop (0.8 mg/kg) was significantly higher in L. casei-treated rats (3.26 microg/mL) than in those untreated rats (2.33 microg/mL) by 40%. Accordingly, the bioavailability of nifedipine was tended to be higher in the former, while the effect of L. casei on the disposition of intravenously administered nifedipine was negligible. We also found that the availability of nifedipine for the passage through the intestinal mucosa was significantly increased in L. casei-treated rats from the single-pass intestinal perfusion experiments. Therefore, it is likely that the exposure to nifedipine after its administration into rat intestine was increased by oral ingestion of L. casei due to an increase in absorption by increased intestinal availability (decreased metabolic extraction) during passage through the intestinal mucosa. This study has suggested that L. casei has some effect on the metabolic activity in the intestinal mucosa, though it seems to be only mild. [Abstract/Link to Full Text]

Chono S, Tanino T, Seki T, Morimoto K
Pharmacokinetic and pharmacodynamic efficacy of intrapulmonary administration of ciprofloxacin for the treatment of respiratory infections.
Drug Metab Pharmacokinet. 2007 Apr;22(2):88-95.
The pharmacokinetic and pharmacodynamic efficacy of intrapulmonary administration of ciprofloxacin (CPFX) for the treatment of respiratory infections caused by pathogenic microorganisms resisting sterilization systems of alveolar macrophages (AMs) was evaluated by comparison with an oral administration. The time-courses of the concentration of CPFX in AMs and lung epithelial lining fluid (ELF) following intrapulmonary administration of CPFX solution to rats (200 microg/kg) were markedly higher than that following oral administration (10 mg/kg). The time-course of the concentrations of CPFX in plasma following intrapulmonary administration was markedly lower than that in AMs and ELF. These results indicate that intrapulmonary administration is more effective in delivering CPFX to AMs and ELF, compared with oral administration, in spite of a low dose and it avoids distribution of CPFX to the blood. In addition, the antibacterial effects of CPFX in AMs and ELF following intrapulmonary administration were evaluated by pharmacokinetics/pharmacodynamics analysis. The concentration of CPFX in AMs and ELF-time curve (AUC)/minimum inhibitory concentration of CPFX (MIC) ratio and the maximum concentration of CPFX in AMs and ELF (Cmax)/MIC ratio were markedly higher than the effective values. The present study indicates that intrapulmonary administration of CPFX is an effective technique for the treatment of respiratory infections. [Abstract/Link to Full Text]

Benetton SA, Fang C, Yang YO, Alok R, Year M, Lin CC, Yeh LT
P450 phenotyping of the metabolism of selegiline to desmethylselegiline and methamphetamine.
Drug Metab Pharmacokinet. 2007 Apr;22(2):78-87.
Although there is evidence in the literature of the participation of CYP2B6 in the metabolism of selegiline, it is not clear which other CYP isoforms contribute to its metabolism. The aim of this study was to investigate the P450 isozymes (CYPs) involved in the metabolism of selegiline to desmethylselegiline (DMS) and methamphetamine (MA) using four assays: incubation of selegiline with cDNA expressed CYPs, inhibition of DMS and MA formations in human liver microsomes by CYP-selective chemical inhibitors or CYP-specific antibodies, and correlation analysis. Correlation analysis, performed in a bank of 15 individual human liver microsomes, yielded correlation coefficients for DMS and MA formation of 0.769 and 0.792, respectively, for CYP2B6 (p<0.0001) and 0.333 and 0.349, respectively, for CYP3A4 (p<0.05). These results were supported by chemical/specific antibody inhibition assays. The results of correlation analysis and chemical inhibition also indicated that CYP2A6 seems to play a small role in the metabolism of selegiline. These findings confirm that CYP2B6 plays a major role in the metabolism of selegiline and also suggest the involvement of CYP3A4 and CYP2A6. [Abstract/Link to Full Text]

Hayashi M, Tomita M
Mechanistic analysis for drug permeation through intestinal membrane.
Drug Metab Pharmacokinet. 2007 Apr;22(2):67-77.
For drug absorption, intestinal drug permeability's through both the paracellular and transcellular routes were analyzed. Absorption enhancers, such as sodium caprate (C10), decanoylcarnitine (DC) and tartaric acid (TA), increased the paracellular permeability of water-soluble, low lipophilic and poorly absorbable drugs by enlargement of tight junction (TJ) adhering to the intercellular portion; that is, expansion of the paracellular routes. C10 increased the intracellular calcium level to induce contraction of calmodulin-dependent actin filaments. Although DC also increased the intracellular calcium level, the action was independent of calmodulin, and thus the action mechanism of DC was considered to differ from that of C10. DC and TA decreased the intracellular ATP level and the intracellular pH, suggesting that intracellular acidosis increases the calcium level through decrease in ATP level followed by opening TJ. TA had no effect on Western blot analysis, but TA significantly inhibited excretion of rhodamine 123, one of the P-glycoprotein (P-gp) substrates, from the serosal to mucosal side, suggesting that TA increases the intestinal absorption of P-gp substrates, possibly by inhibiting the P-gp function without changing the expression of P-gp. During ischemia/reperfusion (I/R) injury during small intestine grafting, TJ opening and decrease in P-gp function simultaneously occurred. The in vitro model of I/R showed that lipid peroxidation is a trigger of the injury, and superoxide and iron ion participate in TJ opening and decrease in P-gp function. Colonic epithelial cells have the specific transcellular transport systems for lipopolysaccharide (LPS), one of which shows substrate specificity in the interaction with CD14 and/or that of TLR4. In the infective disease induced by LPS, the mucosal LPS sensitive transport capability was decreased and in the secretory direction, the receptor-mediated uptake mechanism disappeared. LPS taken up into the cells can be excreted by P-gp or mrp. The expression levels and function of the secretory transporters were considered to be increased in the infective condition. In conclusion, changes in TJ as the membrane structure and P-gp as the membrane function are important factors controlling intestinal membrane transport. [Abstract/Link to Full Text]

Ikeda T
SNP communications: coalition of toxicology and drug metabolism and pharmacokinetics.
Drug Metab Pharmacokinet. 2007 Apr;22(2):65-6. [Abstract/Link to Full Text]

Shimizu M, Tomioka S, Murayama N, Yamazaki H
Missense and nonsense mutations of the flavin-containing monooxygenase 3 gene in a Japanese cohort.
Drug Metab Pharmacokinet. 2007 Feb 25;22(1):61-4.
We sequenced all exons and exon-intron junctions of the flavin-containing monooxygenase 3 (FMO3) gene from 3 Japanese individuals and their family members, who were case subjects that showed low FMO3 metabolic capacity among a population of self-reported trimethylaminuria Japanese volunteers (n=50). We found three novel single nucleotide polymorphisms (SNPs) (g. 20752 A>G, g. 27400 G>A, and g. 30308 C>T) causing an amino acid substitution and stop codons, Asn114Ser in exon 4, Trp388Stop in exon 7, and Gln470Stop in exon 9, respectively. The Trp388Stop and Gln470Stop also presented together with known SNPs, Val257Met and Glu158Lys, respectively, in the same alleles of the FMO3 gene to form novel haplotypes. These sequences are as follows: 1) SNP, 060825Shimizu004; GENE NAME, FMO3; ACCESSION NUMBER, AL021026; LENGTH, 25 base; 5'-TATCCAGTGTAAA/GTAAACATCCTGA-3'. 2) SNP, 060825Shimizu005; GENE NAME, FMO3; ACCESSION NUMBER, AL021026; LENGTH, 25 base; 5'-CCAGTCCCGCTGG/AGCAGCACAAGTA-3'. 3) SNP, 060825Shimizu006; GENE NAME, FMO3; ACCESSION NUMBER, AL021026; LENGTH, 25 base; 5'-TGTAGTCCCTACC/TAGTTTAGGCTGG-3'. [Abstract/Link to Full Text]

Fukushima-Uesaka H, Saito Y, Tohkin M, Maekawa K, Hasegawa R, Kawamoto M, Kamatani N, Suzuki K, Yanagawa T, Kajio H, Kuzuya N, Yasuda K, Sawada J
Genetic variations and haplotype structures of the ABC transporter gene ABCC1 in a Japanese population.
Drug Metab Pharmacokinet. 2007 Feb 25;22(1):48-60.
Multidrug resistance-related protein 1 (MRP1), an ATP-binding cassette transporter encoded by the ABCC1 gene, is expressed in many tissues, and functions as an efflux transporter for glutathione-, glucuronate- and sulfate-conjugates as well as unconjugated substrates. In this study, the 31 exons and their flanking introns of ABCC1 were comprehensively screened for genetic variations in 153 Japanese subjects to elucidate the linkage disequilibrium (LD) profiles and haplotype structures of ABCC1 that is necessary for pharmacogenetic studies of the substrate drugs. Eighty-six genetic variations including 31 novel ones were found: 1 in the 5'-flanking region, 1 in the 5'-untranslated region (UTR), 20 in the coding exons (9 synonymous and 11 nonsynonymous variations), 4 in the 3'-UTR, and 60 in the introns. Of these, eight novel nonsynonymous variations, 726G>T (Trp242Cys), 1199T>C (Ile400Thr), 1967G>C (Ser656Thr), 2530G>A (Gly844Ser), 3490G>A (Val1164Ile), 3550G>A (Glu1184Lys), 3901C>T (Arg1301Cys), and 4502A>G (Asp1501Gly), were detected with an allele frequency of 0.003. Based on the LD profiles, the analyzed regions of the gene were divided into five LD blocks (Blocks -1 and 1 to 4). The multiallelic repeat polymorphism in the 5'-UTR was defined as Block -1. For Blocks 1, 2, 3 and 4, 32, 23, 23 and 13 haplotypes were inferred, and 9, 7, 7 and 6 haplotypes commonly found on > or = 10 chromosomes accounted for > or = 91% of the inferred haplotypes in each block. Haplotype-tagging single nucleotide polymorphisms for each block were identified to capture the common haplotypes. This study would provide fundamental and useful information for the pharmacogenetic studies of MRP1-dependently effluxed drugs in Japanese. [Abstract/Link to Full Text]

Shibayama T, Sugiyama D, Kamiyama E, Tokui T, Hirota T, Ikeda T
Characterization of CS-023 (RO4908463), a novel parenteral carbapenem antibiotic, and meropenem as substrates of human renal transporters.
Drug Metab Pharmacokinet. 2007 Feb 25;22(1):41-7.
To characterize the renal handling of CS-023 (RO4908463), a novel parenteral carbapenem antibiotic, and meropenem in humans, we examined their affinities as substrates to human renal transporters. In vitro studies on the uptake of [14C]CS-023 and [14C]meropenem were conducted using HEK293 cells expressing human organic anion transporters (hOAT) 1, hOAT3, hOAT4, and the human organic cation transporters (hOCT) 1 and hOCT2. CS-023 did not serve as the substrate for any of the transporters tested. On the other hand, meropenem was transported by hOAT1 and hOAT3. The Km value of the hOAT3-mediated transport was 847 microM, and the uptake was inhibited by probenecid, p-aminohippurate and benzylpenicillin with Ki values of 3.76, 712, and 202 microM, respectively. One of the reasons why CS-023 is not a substrate of hOATs, and vice versa for meropenem, would be that a very small proportion of CS-023 exists as the anionic form at the physiological pH, whereas 50% of meropenem exists as the anionic form. These findings indicate that the lack of recognition of CS-023 by renal transporters is one of the reasons for its long plasma half-life in humans compared with meropenem which undergoes renal tubular secretion mediated by hOAT1 and hOAT3. [Abstract/Link to Full Text]

Narawa T, Tsuda Y, Itoh T
Chiral recognition of amethopterin enantiomers by the reduced folate carrier in Caco-2 cells.
Drug Metab Pharmacokinet. 2007 Feb 25;22(1):33-40.
Stereoselectivity of the human reduced folate carrier (RFC1) in Caco-2 cells was examined using methotrexate (L-amethopterin, L-MTX) and its antipode (D-amethopterin, D-MTX) as model substrates. The initial uptake rate of L-MTX into Caco-2 cells followed Michaelis-Menten kinetics with a Km value of approximately 1 microM. The Eadie-Hofstee plot of the RFC1-mediated L-MTX uptake showed that it was mediated by a single transport system, RFC1. Dixon plots revealed that L-MTX uptake was inhibited competitively by folic acid (FA), L-MTX and D-MTX, with Ki values of approximately 0.8, 1.5 and 180 microM, respectively. The results showed that the affinities of FA and L-MTX to RFC1 were approximately 120-fold greater than that of D-MTX. The uptake of L- and D-MTX into Caco-2 cells was also measured using LC-MS/MS analysis, which revealed that the L-MTX uptake was at least 7-fold greater than that of D-MTX. The present study revealed significant stereoselectivity of RFC1 toward amethopterin enantiomers with the L-isomer being much more favored. [Abstract/Link to Full Text]

Fujii Y, Takahashi M, Morita H, Kikuchi H, Aramaki Y, Amidon GL
Characteristics of gastrointestinal absorption of DX-9065a, a new synthetic anticoagulant.
Drug Metab Pharmacokinet. 2007 Feb 25;22(1):26-32.
DX-9065a, a newly synthesized anticoagulant that selectively inhibits factor Xa, is a zwitterion and has characteristics of high water solubility and low lipophilicity. We predicted the fraction absorbed (Fa) of DX-9065a to be approximately 15-35% in humans, based on the boundary layer theory using the intestinal perfusion method in rats. However, human oral bioavailability was 2-3% in clinical trials, and the result of actual human bioavailability was lower than that of the predicted Fa. Thus, in this report, the reason for low oral bioavailability of DX-9065a was examined by in vitro and in vivo experiments. The factors affecting oral bioavailability of DX-9065a were not the hepatic first-pass effect, degradation of the drug in intestinal fluid, nor the interaction of the drug with the intestinal mucin. Furthermore, no effect of P-gp efflux was observed. Oral absorption of the drug in rats with bile duct ligation was significantly higher than that in normal rats with bioavailability of 17 and 3%, respectively. It was confirmed that bile acids inhibited DX-9065a absorption because DX-9065a interacted with bile acids to form insoluble complexes. The results suggest that the complex formation of DX-9065a with bile acids in the intestinal tract is an important factor affecting absorption of DX-9065a. [Abstract/Link to Full Text]

Furuya Y, Ozeki T, Takayanagi R, Yokoyama H, Okuyama K, Yamada Y
Theory based analysis of anti-inflammatory effect of infliximab on Crohn's disease.
Drug Metab Pharmacokinet. 2007 Feb 25;22(1):20-5.
Tumor necrosis factor (TNF)-alpha, a primary mediator of inflammatory responses, is increased in patients with active Crohn's disease (CD) and considered to play an important role in the regulation of inflammation in CD. Infliximab (IFX) is a chimeric murine-human monoclonal IgG1 antibody that targets TNF-alpha and is used as a therapeutic agent for CD. Although that dosage regimen has been established through clinical trial experience, it has not been analyzed theoretically. We analyzed of sequential changes of the Crohn's disease activity index (CDAI) using a pharmacokinetic-pharmacodynamic model integrating the pharmacokinetics of IFX and turnover rate of TNF-alpha. The time course effects of IFX derived from the present model were matched to reported data regarding CDAI ratios, and we found that the clinical effect of IFX reached a maximum value 2 to 4 weeks after administration and was maintained for the next several weeks. Our results suggested that the standard dosage regimen of IFX is theoretically appropriate. Further, based on the results of various dosage regimens, a second administration of IFX 2 weeks after the first dose was shown to achieve remission in the early stage of active CD, when IFX was given as a repeated treatment. [Abstract/Link to Full Text]

Minamiyama Y, Takemura S, Imaoka S, Funae Y, Okada S
Cytochrome P450 is responsible for nitric oxide generation from NO-aspirin and other organic nitrates.
Drug Metab Pharmacokinet. 2007 Feb 25;22(1):15-9.
Nitric oxide (NO) biotransformation from NO-aspirin (NCX-4016) is not clearly understood. We have previously reported that cytochrome P450 (P450) plays important role in NO generation from other organic nitrates such as nitroglycerin (NTG) and isosorbide dinitrate (ISDN). The present study was designed to elucidate the role of human cytochrome P450 isoforms in NO formation from NCX-4016, using lymphoblast microsomes transfected with cDNA of human P450 or yeast-expressed, purified P450 isoforms. CYP1A2 and CYP2J2, among other isoforms, were strongly related to NO production from NCX-4016. In fact, these isoforms were detected in human coronary endothelial cells. These results suggest that NADPH-cytochrome P450 reductase and the P450 system participate in NO formation from NCX-4016, as well as other organic nitrates. [Abstract/Link to Full Text]

Ohdo S
Chronopharmacology focused on biological clock.
Drug Metab Pharmacokinet. 2007 Feb 25;22(1):3-14.
The mammalians circadian pacemaker resides in the paired suprachiasmatic nuclei (SCN) and influences a multitude of biological processes, including the sleep-wake rhythm. Clock genes are the genes that control the circadian rhythms in physiology and behavior. The effectiveness and toxicity of many drugs vary depending on dosing time associated with 24 hr rhythms of biochemical, physiological and behavioral processes under the control of circadian clock. Such chronopharmacological phenomena are influenced by not only the pharmacokinetics but also pharmacodynamics of medications. Identification of a rhythmic marker for selecting dosing time will lead to improved progress and diffusion of chronopharmacotherapy. The mechanisms underlying chronopharmacological findings should be clarified from the viewpoint of clock genes. On the other hand, several drugs have an effect on circadian clock. The knowledge of interactions between circadian clock and drugs should be very useful for clinical practice. Therefore, I introduce the regulatory system of biological rhythm from viewpoints of clock genes and the possibility of pharmacotherapy based on clock genes. [Abstract/Link to Full Text]

Kim SR, Ozawa S, Saito Y, Kurose K, Kaniwa N, Kamatani N, Hamaguchi T, Shirao K, Muto M, Ohtsu A, Yoshida T, Matsumura Y, Saijo N, Sawada J
Fourteen novel genetic variations and haplotype structures of the TYMS gene encoding human thymidylate synthase (TS).
Drug Metab Pharmacokinet. 2006 Dec;21(6):509-16.
Forty genetic variations including 14 novel ones were found in the human TYMS gene, which encodes thymidylate synthase, in 263 Japanese cancer patients who received 5-fluorouracil (FU)-based chemotherapy. Three novel variations were located within the 28-bp tandem repeat sequence in the 5'-untranslated region (UTR) and were designated 5Rc, 3Rc-ins and 4Rc. Allele frequencies were 0.021 for 5Rc, 0.006 for 3Rc-ins and 0.002 for 4Rc. Other novel variations included -133G>C and -125G>C in the 5'-UTR; IVS1-278G>A, IVS2-68C>T, IVS2-23T>C, IVS4+122_+123insATTG, IVS4-141G>A, IVS5-100A>T and IVS6-111G>A in the introns; and 1244(*302)A>G and 1264(*322)G>A in the 3'-UTR. The allele frequencies were 0.34 for IVS4+122_+123insATTG, 0.042 for -133G>C, 0.011 for IVS4-141G>A, 0.006 for -125G>C, 0.004 for IVS1-278G>A, IVS2-68C>T, 1244(*302)A>G and 1264(*322)G>A, and 0.002 for IVS2-23T>C, IVS5-100A>T and IVS6-111G>A. Using the detected polymorphisms, linkage disequilibrium (LD) analysis was performed, which divided the TYMS gene into three LD blocks. The 28-bp tandem repeat sequence in the 5'-UTR was assigned as Block 2 with a total of 7 alleles. In Blocks 1 and 3, 7 and 19 haplotypes were determined/inferred, respectively. Our findings provide fundamental and useful information for genotyping TYMS in the Japanese and probably other Asian populations. [Abstract/Link to Full Text]

Ramasamy K, Sisy Sam S, Chandrasekaran A
Allele and genotype frequency of MDR1 C3435T in Tamilian population.
Drug Metab Pharmacokinet. 2006 Dec;21(6):506-8.
The allele and genotype frequencies of MDR1 C3435T polymorphism were determined in 185 unrelated healthy Tamilians. The genomic DNA was extracted from peripheral leucocytes using phenol chloroform method and genotyped by PCR-RFLP method. The frequencies of MDR1 C3435 and T3435 alleles in Tamilian population were 0.46 and 0.54 respectively. The distribution of T3435 in this population was found to be greater than Africans and almost similar to Caucasians and Orientals. The distribution of CC, CT and TT genotypes was 0.18, 0.56 and 0.26 respectively. The frequency distribution of the CC genotype was lower in them when compared with Chinese and Africans whereas CT genotype was higher in comparison with all the major ethnic groups. [Abstract/Link to Full Text]

Fukumoto K, Kobayashi T, Tachibana K, Kato R, Tanaka K, Komamura K, Kamakura S, Kitakaze M, Ueno K
Effect of amiodarone on the serum concentration/dose ratio of metoprolol in patients with cardiac arrhythmia.
Drug Metab Pharmacokinet. 2006 Dec;21(6):501-5.
Amiodarone has pharmacokinetic interactions with a number of therapeutic drugs, including warfarin, phenytoin, flecainide, and cyclosporine. Metoprolol is mainly metabolized by CYP2D6, and desethylamiodarone, a metabolite of amiodarone, has a markedly greater inhibitory effect on CYP2D6 than amiodarone. Therefore, the goal of this study was to evaluate the effect of amiodarone and desethylamiodarone on the serum concentration/dose ratio (C/D) of metoprolol in 120 inpatients with cardiac arrhythmias that received either metoprolol and amiodarone (MET+AMD group, n=30) or metoprolol alone (MET group, n=90). The ratio of administered metoprolol was compared between the MET and the MET+AMD groups. The dose of metoprolol and patient age were significantly higher in the MET group when compared with the MET+AMD group (1.00+/-0.480 versus 0.767+/-0.418 mg/kg/day, p<0.050; 68.6+/-10.6 versus 57.6+/-14.1 years, p<0.001, respectively), but the C/D ratio was significantly lower in the MET group than in the MET+AMD group (90.8+/-64.0 versus 136+/-97.8, p<0.01). Furthermore, a significant correlation was found between the C/D ratio and desethylamiodarone concentration (n=30, r=0.371, p<0.01). The results suggest that there is a significant interaction between amiodarone and metoprolol via desethylamiodarone-induced inhibition of CYP2D6. Therefore, careful monitoring of metoprolol concentrations/bioactivity of CYP2D6 is required in the context of co-administration of amiodarone and metoprolol. [Abstract/Link to Full Text]

Sato E, Shimomura M, Masuda S, Yano I, Katsura T, Matsumoto S, Okitsu T, Iwanaga Y, Noguchi H, Nagata H, Yonekawa Y, Inui K
Temporal decline in sirolimus elimination immediately after pancreatic islet transplantation.
Drug Metab Pharmacokinet. 2006 Dec;21(6):492-500.
Pancreatic islet transplantation is a curable treatment for type 1 diabetes and has been put into practice in various countries. In this study, we analyzed the pharmacokinetic characteristics of sirolimus and tacrolimus in six Japanese patients with pancreatic islet transplants immediately after surgery, and monitored efficacy and toxicity. The patients were treated with immunosuppressive therapy based on the Edmonton protocol, that is, sirolimus and low-dose tacrolimus. Pharmacokinetic analyses were performed using the nonlinear mixed-effects modeling program NONMEM. Large inter- and intra-individual variability was observed in the pharmacokinetics of sirolimus and tacrolimus. A model with increased apparent clearance in the postoperative period explained well the intra-individual variability in the pharmacokinetics of both drugs. The most frequent drug-induced toxicity was a decrease in the white blood cell count, and two of six patients required the administration of granulocyte colony-stimulating factor. Clinical laboratory tests immediately before the transplantation and cytochrome P450 3A5 genotype were not related to the high blood concentrations of sirolimus after the loading dose. From these results, the apparent clearance of sirolimus and tacrolimus might temporally decline immediately after pancreatic islet transplantation. A high trough concentration of sirolimus might increase the risk of hematological toxicy, and adjustment of the dosage for immunosuppressive treatment will be necessary in Japanese patients. [Abstract/Link to Full Text]

Moriyasu A, Sugihara K, Nakatani K, Ohta S, Kitamura S
In vivo-in vitro relationship of methotrexate 7-hydroxylation by aldehyde oxidase in four different strain rats.
Drug Metab Pharmacokinet. 2006 Dec;21(6):485-91.
The in vivo metabolism of methotrexate (MTX) to 7-hydroxymethotrexate (7-OH-MTX) was studied using four strains of rats. When MTX was administered to these rats, 7-OH-MTX was detected as the major in vivo metabolite, mainly in bile and feces, and also slightly in the urine. There were marked strain differences in the amounts of 7-OH-MTX excreted in bile, feces and urine. The highest recovery of 7-OH-MTX in bile, feces and urine was observed in Sea:SD rats (6.2%, 4.2% and 0.8% of dose, respectively), followed by Jcl:SD and Crj:SD rats. The lowest recovery (0.02%, 0.2% and 0.003%, respectively) was observed in WKA/Sea rats. The variations of excreted amount of 7-OH-MTX were closely correlated with the strain differences of cytosolic MTX 7-hydroxylase and benzaldehyde oxidase activities. Our results indicate that variation of formation of 7-OH-MTX from MTX in vivo in rats is due primarily to variation of aldehyde oxidase. [Abstract/Link to Full Text]

Peamkrasatam S, Sriwatanakul K, Kiyotani K, Fujieda M, Yamazaki H, Kamataki T, Yoovathaworn K
In vivo evaluation of coumarin and nicotine as probe drugs to predict the metabolic capacity of CYP2A6 due to genetic polymorphism in Thais.
Drug Metab Pharmacokinet. 2006 Dec;21(6):475-84.
The association between the distribution characteristics of CYP2A6 catalytic activities toward nicotine and coumarin, and the frequency distribution of CYP2A6 variant alleles reported was estimated in 120 healthy Thais. The distributions of the subjects as classified by the amounts of 7-hydroxycoumarin (7-OHC) excreted in the urine and by cotinine/nicotine ratio in the plasma were clearly bimodal. However, the numbers of apparently poor metabolizers for coumarin and nicotine were different. The inter-individual variability in the in vivo dispositions of coumarin and nicotine closely related to the CYP2A6 genetic polymorphism. There was a close correlation between the rate of 7-OHC excretion in the urine and cotinine/nicotine ratio in the plasma among subjects (R=0.92, p<0.001). The frequency of CYP2A6 allele found in the present study was: CYP2A6*1A=32% (95% CI, 22.1-39.4%), CYP2A6*1B=27% (95% CI, 19.4-33.5%), CYP2A6*9=20% (95% CI, 17.6-23.3%), CYP2A6*4=14% (95% CI, 9.6-17.8%), CYP2A6*7=5% (95% CI, 3.7-9.4%), CYP2A6*10=2% (95% CI, 0.8-5.1%). Subjects having CYP2A6*1A/*1B were found to have a higher rate of 7-OHC excretion, as well as a higher cotinine/nicotine ratio in the plasma compared with those of the other genotypes. In contrast, subjects with CYP2A6*4/*7 and CYP2A6*7/*7 almost lacked any cotinine formation, whereas urinary 7-OHC was still detectable. CYP2A6*9 allele clearly resulted in reduced enzyme activities. Despite the absence of the homozygote for CYP2A6*10 allele, the presence of CYP2A6*10 allele significantly decreased the enzyme activities. The results of the present study demonstrate that in vivo phenotyping of CYP2A6 using nicotine and coumarin are not metabolically equivalent. Nicotine is a better probe according to its specificity, while coumarin is still valuable to be used for a routine CYP2A6 phenotyping since the test employs a non-invasive method. [Abstract/Link to Full Text]

Yoshitsugu H, Nishimura M, Tateno C, Kataoka M, Takahashi E, Soeno Y, Yoshizato K, Yokoi T, Naito S
Evaluation of human CYP1A2 and CYP3A4 mRNA expression in hepatocytes from chimeric mice with humanized liver.
Drug Metab Pharmacokinet. 2006 Dec;21(6):465-74.
We investigated and compared the expression of human CYPs mRNA in primary cultures of cryopreserved human hepatocytes and in chimeric mice constructed by transplanting hepatocytes from the same human donors. Analysis was performed by real-time reverse-transcription polymerase chain reaction. Initial expression levels for the 12 human CYPs mRNA in chimeric mouse hepatocytes were higher than those in human hepatocytes, but a low correlation coefficient was observed (r=0.690). After 24 h of culture, the correlation remained low (r=0.699). The medium was replaced with fresh medium without human epidermal growth factor, and after 48 h of culture, expression of the 12 human CYPs mRNA were very similar in human hepatocytes and chimeric mouse hepatocytes, and a higher correlation coefficient was observed (r=0.809). After 72 h of culture, the correlation remained high (r=0.873). The ratio of human CYP1A2 mRNA to beta-actin mRNA in chimeric mouse hepatocytes decreased quickly during the first 24 h of culture, and then remained constant. Expression profiles of human CYP1A2 mRNA in chimeric mouse hepatocytes were similar to those in human hepatocytes after exposure of beta-naphthoflavone. CYP3A4 mRNA expression was increased significantly by rifampicin (Rif) exposure in human hepatocytes, whereas Rif-induced increases in CYP3A4 mRNA expression in chimeric mouse hepatocytes was seen for two of the three donors. In conclusion, we demonstrated that expression and induction of human CYPs in human hepatocytes can be reproduced in chimeric mouse hepatocytes. [Abstract/Link to Full Text]

Takemoto S, Yamaoka K, Nishikawa M, Takakura Y
Histogram analysis of pharmacokinetic parameters by bootstrap resampling from one-point sampling data in animal experiments.
Drug Metab Pharmacokinet. 2006 Dec;21(6):458-64.
A bootstrap method is proposed for assessing statistical histograms of pharmacokinetic parameters (AUC, MRT, CL and V(ss)) from one-point sampling data in animal experiments. A computer program, MOMENT(BS), written in Visual Basic on Microsoft Excel, was developed for the bootstrap calculation and the construction of histograms. MOMENT(BS) was applied to one-point sampling data of the blood concentration of three physiologically active proteins ((111)In labeled Hsp70, Suc(20)-BSA and Suc(40)-BSA) administered in different doses to mice. The histograms of AUC, MRT, CL and V(ss) were close to a normal (Gaussian) distribution with the bootstrap resampling number (200), or more, considering the skewness and kurtosis of the histograms. A good agreement of means and SD was obtained between the bootstrap and Bailer's approaches. The hypothesis test based on the normal distribution clearly demonstrated that the disposition of (111)In-Hsp70 and Suc(20)-BSA was almost independent of dose, whereas that of (111)In-Suc(40)-BSA was definitely dose-dependent. In conclusion, the bootstrap method was found to be an efficient method for assessing the histogram of pharmacokinetic parameters of blood or tissue disposition data by one-point sampling. [Abstract/Link to Full Text]

Nakata K, Tanaka Y, Nakano T, Adachi T, Tanaka H, Kaminuma T, Ishikawa T
Nuclear receptor-mediated transcriptional regulation in Phase I, II, and III xenobiotic metabolizing systems.
Drug Metab Pharmacokinet. 2006 Dec;21(6):437-57.
Studies of the genetic regulation involved in drug metabolizing enzymes and drug transporters are of great interest to understand the molecular mechanisms of drug response and toxic events. Recent reports have revealed that hydrophobic ligands and several nuclear receptors are involved in the induction or down-regulation of various enzymes and transporters involved in Phase I, II, and III xenobiotic metabolizing systems. Nuclear receptors (NRs) form a family of ligand-activated transcription factors (TFs). These proteins modulate the regulation of target genes by contacting their promoter or enhancer sequences at specific recognition sites. These target genes include metabolizing enzymes such as cytochrome P450s (CYPs), transporters, and NRs. Thus it was now recognized that these NRs play essential role in sensing processing xenobiotic substances including drugs, environmental chemical pollutants and nutritional ingredients. From literature, we picked up target genes of each NR in xenobiotic response systems. Possible cross-talk, by which xenobiotics may exert undesirable effects, was listed. For example, the role of NRs was comprehensively drawn up in cholesterol and bile acid homeostasis in human hepatocyte. Summarizing current states of related research, especially for in silico response element search, we tried to elucidate nuclear receptor mediated xenobiotic processing loops and direct future research. [Abstract/Link to Full Text]

Okuda M, Kimura N, Inui K
Interactions of fluoroquinolone antibacterials, DX-619 and levofloxacin, with creatinine transport by renal organic cation transporter hOCT2.
Drug Metab Pharmacokinet. 2006 Oct;21(5):432-6.
Interactions of DX-619, a novel fluoroquinolone antibacterial, and levofloxacin (LVFX) with the human renal organic cation transporter hOCT2 were studied. The intracellular accumulation of [(14)C]creatinine in stable transfectants of HEK293 cells expressing hOCT2 (hOCT2-HEK293) as well as vector-transfected HEK293 cells (VEC-HEK293) was evaluated in the presence of DX-619 and LVFX at various concentrations. When added extracellularly, both DX-619 and LVFX inhibited the uptake of [(14)C]creatinine (5 microM) by hOCT2-HEK293 cells in a dose-dependent manner. Unlike in hOCT2-HEK293 cells, the uptake in VEC-HEK293 cells was not inhibited by either fluoroquinolone suggesting that hOCT2 was specifically involved in the inhibition. The apparent IC(50) value for the inhibition of [(14)C]creatinine uptake in hOCT2-HEK293 cells was 1.29+/-0.23 microM for DX-619 and 127+/-27 microM for LVFX, indicating DX-619 to be approximately 100-fold more potent than LVFX at inhibiting the transport of [(14)C]creatinine by hOCT2. A Dixon plot revealed that the inhibition by DX-619 of the hOCT2-mediated transport of [(14)C]creatinine was competitive. Fluoroquinolone antibacterials have the ability to inhibit the transport of creatinine by hOCT2, with DX-619 being much more effective than LVFX. [Abstract/Link to Full Text]

Recent Articles in Journal of Pharmacological Sciences

Kim DH, Yoon BH, Kim YW, Lee S, Shin BY, Jung JW, Kim HJ, Lee YS, Choi JS, Kim SY, Lee KT, Ryu JH
The seed extract of Cassia obtusifolia ameliorates learning and memory impairments induced by scopolamine or transient cerebral hypoperfusion in mice.
J Pharmacol Sci. 2007 Sep;105(1):82-93.
In the present study, we assessed the effect of the ethanolic extract of the seeds of Cassia obtusifolia (COE) on the learning and memory impairments induced by scopolamine or transient bilateral common carotid artery occlusion (2VO). In a study of the cholinergic dysfunction induced by scopolamine, single COE (25, 50, or 100 mg/kg, p.o.) administration significantly attenuated scopolamine-induced cognitive impairments as determined by the passive avoidance and Y-maze tasks (P<0.05) and also reduced escape-latency on the Morris water maze task (P<0.05). In the 2VO study, COE (50 mg/kg, p.o.) significantly reversed 2VO-induced cognitive impairments in mice by the passive avoidance and the Y-maze tasks (P<0.05). Moreover, COE (50 mg/kg, p.o.) also reduced escape-latency and prolonged swimming time in the target quadrant during a probe trial of the Morris water maze task (P<0.05). In an in vitro study, COE was found to inhibit acetylcholinesterase activity in a dose-dependent manner (IC(50) value: 81.6 microg/ml). Furthermore, COE also inhibited acetylcholinesterase activity in an ex vivo study. These results suggest that COE attenuates memory impairment induced by scopolamine or 2VO and that these effects are mediated by enhancing the cholinergic nervous system via acetylcholinesterase inhibition. [Abstract/Link to Full Text]

Lee HM, Lee CK, Lee SH, Roh HY, Bae YM, Lee KY, Lim J, Park PJ, Park TK, Lee YL, Won KJ, Kim B
p38 mitogen-activated protein kinase contributes to angiotensin II-stimulated migration of rat aortic smooth muscle cells.
J Pharmacol Sci. 2007 Sep;105(1):74-81.
In this study, we clarified the intracellular mechanism of angiotensin II (Ang II) in promoting migration in rat aortic smooth muscle cells (RASMCs). RASMC migration was measured with the Boyden chamber assay, and the result was confirmed with an aortic sprout assay. The activities of kinases were investigated by western blot analysis. Ang II enhanced RASMC migration, which was chemotaxis directed, and induced the phosphorylation of p38 mitogen-activated protein kinase (MAPK), extracellular signal-regulated kinase 1/2 (ERK1/2), and heat shock protein 27 (Hsp27). Ang II-enhanced cell migration was inhibited by SB203580 (a p38 MAPK inhibitor) and piceatannol (a spleen tyrosine kinase inhibitor), but only partially by PD98059 (an ERK inhibitor) and PP2 (a Src inhibitor). The Ang II-stimulated phosphorylation of p38 MAPK and Hsp27 in RASMCs was inhibited by piceatannol and SB203580. The phosphorylation of ERK1/2 stimulated by Ang II was suppressed by PD98059, piceatannol, and PP2. Ang II increased the sprout outgrowth from aortic rings and this response was attenuated by pretreatment with SB203580, PD98059, PP2, or piceatannol. These results suggest that p38 MAPK contributes to the regulation of the Ang II-induced chemotactic migration of vascular smooth muscle cells, which is mediated by Hsp27 phosphorylation. [Abstract/Link to Full Text]

Hegyesi H, Colombo L, Pállinger E, Tóth S, Boer K, Molnár V, Falus A
Impact of systemic histamine deficiency on the crosstalk between mammary adenocarcinoma and T cells.
J Pharmacol Sci. 2007 Sep;105(1):66-73.
The purpose of the present study was to investigate the influence of lack of histamine (HA) on tumor growth and functions of T cells in order further to illustrate the mechanism of immunological tolerance induction by HA. We assessed the phenotype and cytokine production of splenic lymphocytes in syngeneic HA-free (histidine decarboxylase knock-out) (HDC KO) and wild-type mice, inoculated subcutaneously with the LM2 murine breast cancer cell line. Relative quantification of target mRNA was performed with a TaqMan real-time RT-PCR assay. The CD4(+)CD25(high+) Treg cell numbers were significantly smaller in the tumor-bearing KO mice than in the wild type ones measured by flow-cytometry. The expression of forkhead box P3 (Foxp3) decreased significantly and the copies of splenic Tbox-21 (T-bet) transcriptional factor mRNA was higher in HDC KO tumor-bearing mice than those of normal mice. The cytokine levels showed that a smaller number of interleukin-13-producing Th2 cells were elicited compared to interferon-gamma-producing Th1 cells in the tumor-bearing HDC KO mice. In conclusion, the present study demonstrates that endogenous histamine stimulates the growth of breast adenocarcinoma tumor implants in mice by suppressing anti-tumor immunity. [Abstract/Link to Full Text]

Li TJ, Qiu Y, Mao JQ, Yang PY, Rui YC, Chen WS
Protective effects of Guizhi-Fuling-Capsules on rat brain ischemia/reperfusion injury.
J Pharmacol Sci. 2007 Sep;105(1):34-40.
Previous studies revealed that Guizhi-Fuling-Capsules (GZFLC), a traditional Chinese medical (Kampo) formulation composed of five kinds of medicinal plants, Cinnamomum cassia BLUME (Cinnamomi Cortex), Paeonia lactiflora PALL. (Peonies Radix), Paeonia suffruticosa ANDREWS (Moutan Cortex), Prunus persica BATSCH (Persicae Semen), and Poria cocos WOLF (Hoelen), exerts a protective effect against vascular injury and has a protective effect against glutamate- or nitro oxide-mediated neuronal damage. In the present study, the effect of GZFLC in a rat in vivo model of focal cerebral ischemia and reperfusion was investigated. Administration of GZFLC (0.3 and 0.9 g/kg, p.o.) after focal cerebral ischemia significantly decreased brain infarction and water contents in rats subjected to 2-h ischemia followed by 24-h reperfusion from 31.72 +/- 2.49%, 84.76 +/- 1.63% in the model group to 17.31 +/- 3.66%, 82.51 +/- 1.36% and 8.30 +/- 3.73%, 81.35 +/- 1.73%, respectively. Furthermore, analysis of inflammatory cytokines in ischemic brain showed that GZFLC treatment significantly down-regulated expressions of pro-inflammatory cytokines including interleukin (IL)-1beta and tissue necrosis factor-alpha and markedly up-regulated expressions of anti-inflammatory cytokines IL-10 and IL-10R both in mRNA and protein levels. The serum levels of these inflammatory cytokines were also regulated the same way. These results suggested that GZFLC may be beneficial for the treatment of brain ischemia-reperfusion injury partly due to its anti-inflammatory properties. [Abstract/Link to Full Text]

Li D, Cui Q, Chen SG, Wu LJ, Tashiro S, Onodera S, Ikejima T
Inactivation of ras and changes of mitochondrial membrane potential contribute to oridonin-induced autophagy in a431 cells.
J Pharmacol Sci. 2007 Sep;105(1):22-33.
We have previously shown that oridonin isolated from Rabdosia rubescens augmented apoptosis while inhibiting autophagy within 24 h in HeLa cells. However, the mechanisms between apoptosis and autophagy induced by oridonin in A431 cells are largely unknown. Here, it was found that autophagic level is significantly upregulated when A431 cells are pretreated with manumycin A (Ras specific inhibitor) compared with oridonin alone treatment, whereas cells precultured with GW5074 (Raf inhibitor) or PD98059 (ERK inhibitor) did not exhibit such an effect. Ras, but not Raf or ERK, was engaged in the control of oridonin-induced autophagy. At the same time, manumycin A contributes to oridonin-induced downregulation of Ras protein expression. Treatment with the combination of oridonin and manumycin A downregulated phosphorylation of Akt, downstream of phosphatidylinositol 3-OH kinase (PI3-K). Preincubation with the PI3-K inhibitor wortmannin and Akt inhibitor KP372-1 enhanced oridonin-induced apoptosis, whereas it inhibited oridonin-induced autophagy. However, under oridonin treatment, the expression of Beclin-1, which has autophagy-inducing activity, was reduced, suggesting that Beclin-1 did not participate in the oridonin-induced autophagy. Morphologic observations, DNA fragmentation analysis, and LDH activity-based assay showed that 3-methyladenine (3-MA), an inhibitor of autophagy, increased the apoptotic sensitivity of A431 cells to oridonin. In addition, manumycin A contributed to oridonin-induced decrease of mitochondrial membrane potential (Deltapsim), consistent with the upregulation of Bax/Bcl-2 ratio. In conclusion, Ras negatively regulated autophagy in oridonin-treated A431 cells, which might be associated with activation of class I PI3-K. Downregulation of Deltapsim and increasing of the ratio of Bax/Bcl-2 might also be partially responsible for the initiation of the autophagic process. [Abstract/Link to Full Text]

Yadav H, Jain S, Prasad GB, Yadav M
Preventive effect of diabegon, a polyherbal preparation, during progression of diabetes induced by high-fructose feeding in rats.
J Pharmacol Sci. 2007 Sep;105(1):12-21.
In the present study, the polyherbal preparation diabegon, containing 18 plant extracts with hypoglycemic activity, was evaluated for its preventive effect during progression of type 2 diabetes in high-fructose-diet-fed rats. Oral administration of diabegon (100 mg/kg body weight) delayed development of glucose intolerance for 4 weeks in comparison with the diabetic control group, and the effect of diabegon was compared to that of the standard insulin sensitizer drug rosiglitazone. Diabegon treatment also ameliorated the elevation of glycosylated haemoglobin, liver glycogen content, plasma insulin, homeostasis model assessment, free fatty acids, triglycerides, total cholesterol, LDL-cholesterol, and VLDL-cholesterol, whereas it increased HDL-cholesterol after 56 days of treatment (P<0.05). The mechanism of action by which diabegon attenuates insulin resistance and dyslipidemia may be through induction of peroxisome proliferator-activated receptor-gamma and lipoprotein lipase activity in peripheral tissues (muscles). Moreover, diabegon administration for 56 days also produced no alteration in liver and kidney function tests, which seems to indicate its non-toxicity during treatment. Our present results suggest that diabegon may be included in diabetes mellitus treatment regimens, as a drug with good antidiabetic actions but no toxic manifestations. [Abstract/Link to Full Text]

Takahashi S, Jin XL, Kosaka K, Yoshikawa M, Kobayashi H, Oka T
The enhancing effects of peptidase inhibitors on the antinociceptive action of [Met5]enkephalin-Arg6-Phe7 in rats.
J Pharmacol Sci. 2007 Sep;105(1):117-21.
Previous in vitro studies have shown that the degradation of [Met(5)]enkephalin-Arg(6)-Phe(7) during incubation with cerebral membrane preparations is largely prevented by a mixture of three peptidase inhibitors: amastatin, captopril, and phosphoramidon. The present in vivo study shows that the inhibitory effect of [Met(5)]enkephalin-Arg(6)-Phe(7) administered intra-third-ventricularly on the tail-flick response was increased more than 1000-fold by the intra-third-ventricular pretreatment with three peptidase inhibitors. The antinociceptive effect produced by the [Met(5)]enkephalin-Arg(6)-Phe(7) in rats pretreated with any combination of two peptidase inhibitors was significantly smaller than that in rats pretreated with three peptidase inhibitors, indicating that any residual single peptidase could inactivate significant amounts of the [Met(5)]enkephalin-Arg(6)-Phe(7). The present data, together with those obtained from previous studies, clearly show that amastatin-, captopril-, and phosphoramidon-sensitive enzymes play important roles in the inactivation of endogenous opioid peptides, such as [Met(5)]enkephalin, [Met(5)]enkephalin-Arg(6)-Phe(7), [Met(5)]enkephalin-Arg(6)-Gly(7)-Leu(8), and dynorphin A (1-8), administered intra-third-ventricularly to rats. [Abstract/Link to Full Text]

Tanaka A, Suemaru K, Araki H
A new approach for evaluating renal function and its practical application.
J Pharmacol Sci. 2007 Sep;105(1):1-5.
In clinical practice, the measurement of endogenous serum substances in order to estimate glomerular filtration rate (GFR) is commonly performed, and the serum creatinine level has become the most commonly used serum marker of renal function. However, the measurement of the serum creatinine concentration can sometimes lead to an overestimation of GFR, especially in the elderly. In recent years, it has been suggested that GFR can be predicted based on the serum cystatin C concentrations and that the serum cystatin C concentration is not influenced by gender or age. A recent meta-analysis demonstrated that serum cystatin C is a better marker for GFR than serum creatinine. In clinical practice, it has been suggested that serum cystatin C can optimize early detection for diabetic or hypertensive nephropathy. In addition, the use of serum cystatin C is possibly more appropriate for establishing an appropriate dose adjustment of drugs that are mainly eliminated by the kidney. [Abstract/Link to Full Text]

Ishikawa R, Kohama K
Actin-binding proteins in nerve cell growth cones.
J Pharmacol Sci. 2007 Sep;105(1):6-11.
The motility of the growth cone, an intracellular apparatus located at the tip of the axon in developing neurons, is thought to govern axonal path-finding and the construction of neuronal networks. Growth cones contain an actin-rich cytoskeleton, and their dynamics are regulated by a wide variety of actin-binding proteins and motor proteins. In this review, we will focus on the principal functions of these proteins, their mutual interactions in vitro, and their possible roles in the dynamics of nerve cell growth cones. [Abstract/Link to Full Text]

Kuraishi Y, Ohtsuka E, Nakano T, Kawai S, Andoh T, Nojima H, Kamimura K
Possible involvement of 5-lipoxygenase metabolite in itch-associated response of mosquito allergy in mice.
J Pharmacol Sci. 2007 Sep;105(1):41-7.
This study investigated endogenous mediators involved in mosquito allergy-associated itching in mice. An intradermal injection of an extract of mosquito salivary gland elicited marked scratching in sensitized mice. The 5-lipoxygenase inhibitor zileuton (100 mg/kg), the 5-lipoxygenase activating peptide inhibitor MK-886 (10 mg/kg), and the glucocorticoid betamethasone 17-valerate (3 mg/kg) inhibited the scratching. The scratching was not affected by the cyclooxygenase inhibitors indomethacin and ketoprofen, the TP prostanoid receptor antagonist SQ-29548, the leukotriene B(4) antagonist ONO-4057, the cysteinyl leukotriene antagonist pranlucast, the leukotriene D(4) antagonist MK-571, the platelet-activating factor antagonist CV-3988, the nitric oxide synthase inhibitor N(G)-nitro-L-arginine methyl ester, the H(2) histamine-receptor antagonist cimetidine, the H(1) histamine-receptor antagonist terfenadine plus cimetidine, and cypoheptadine that blocks the 5-HT(1/2) serotonin receptors. Zileuton (100 mg/kg) inhibited the increased activity of the cutaneous nerve branch induced by an intradermal injection of the extract, suggesting the peripheral action. Zileuton and MK-886 (10 and 100 microM) did not affect high K(+)-induced increase in intracellular Ca(2+) concentration in cultured dorsal root ganglion neurons. The results suggest that 5-lipoxygenase metabolite(s) other than leukotriene B(4) and cysteinyl leukotrienes are involved in mosquito allergy-associated itching. [Abstract/Link to Full Text]

Nagate T, Tamura T, Sato F, Kuroda J, Nakayama J, Shibata N
Tranilast suppresses the disease development of the adjuvant- and streptococcal cell wall-induced arthritis in rats.
J Pharmacol Sci. 2007 Sep;105(1):48-56.
This study explores the effects of the anti-allergic and anti-fibrotic agent tranilast on adjuvant- and streptococcal cell wall-induced arthritis in rats, animal models of rheumatoid arthritis in humans. Tranilast (150 or 300 mg/kg, twice daily) or vehicle only was administered orally to the two arthritis models, from 17 days before sensitization. As a comparative control, methotrexate (0.1 mg/kg, once daily) was given to another group. Tranilast suppressed the increase in foot volumes, paw thicknesses, clinical scores, and histopathological scores of the ankle joints in both models dose-dependently. In addition, the fibrosis indices of the ankles were dramatically decreased by tranilast in both of the models. Compared to the effects of methotrexate, tranilast seemed to work more effectively in the streptococcal cell wall-induced arthritis model than in the adjuvant-induced arthritis model. From these observations, it can be concluded that tranilast suppresses the development of arthritis in multiple models and is potentially a novel therapeutic agent for human rheumatoid arthritis. [Abstract/Link to Full Text]

Zhao Y, Migita K, Sato C, Usune S, Iwamoto T, Katsuragi T
Endoplasmic reticulum is a key organella in bradykinin-triggered ATP release from cultured smooth muscle cells.
J Pharmacol Sci. 2007 Sep;105(1):57-65.
ATP has broad functions as an autocrine/paracrine molecule. The mode of ATP release and its intracellular source, however, are little understood. Here we show that bradykinin via B(2)-receptor stimulation induces the extracellular release of ATP via the inositol 1,4,5-trisphosphate [Ins(1,4,5)P(3)]-signaling pathway in cultured taenia coli smooth muscle cells. It was found that bradykinin also increased the production of Ins(1,4,5)P(3) and 2-APB-inhibitable [Ca(2+)](i). The evoked release of ATP was suppressed by the Ca(2+)-channel blockers, nifedipine, and verapamil. Moreover, the extracellular release of ATP was elicited by photoliberation of Ins(1,4,5)P(3). Bradykinin caused a quick and transient accumulation of intracellular ATP from cells treated with 1% perchloric acid solution (PCA), but not with the cell lysis buffer. Peak accumulation was prevented by 2-APB and thapsigargin, but not by nifedipine or verapamil, inhibitors of extracellular release of ATP. These findings suggest that bradykinin elicits the extracellular release of ATP that is mediated by the Ins(1,4,5)P(3)-induced Ca(2+) signaling and, finally, leads to a Ca(2+)-dependent export of ATP from the cells. Furthermore, the bradykinin-induced transient accumulation of ATP in the cells treated with PCA may imply a possible release of ATP from the endoplasmic reticulum. [Abstract/Link to Full Text]

Horinouchi T, Miyake Y, Nishiya T, Nishimoto A, Yorozu S, Jinno A, Kajita E, Miwa S
Characterization of noradrenaline-induced increases in intracellular Ca2+ levels in Chinese hamster ovary cells stably expressing human alpha1A-adrenoceptor.
J Pharmacol Sci. 2007 Sep;105(1):103-11.
The mechanism for noradrenaline (NA)-induced increases in intracellular Ca(2+) concentration ([Ca(2+)](i)) and physiological significance of Na(+) influx through receptor-operated channels (ROCs) and store-operated channels (SOCs) were studied in Chinese hamster ovary (CHO) cells stably expressing human alpha(1A)-adrenoceptor (alpha(1A)-AR). [Ca(2+)](i) was measured using the Ca(2+) indicator fura-2. NA (1 microM) elicited transient and subsequent sustained [Ca(2+)](i) increases, which were inhibited by YM-254890 (G(alphaq/11) inhibitor), U-73122 (phospholipase C (PLC) inhibitor), and bisindolylmaleimide I (protein kinase C (PKC) inhibitor), suggesting their dependence on G(alphaq/11)/PLC/PKC. Both phases were suppressed by extracellular Ca(2+) removal, SK&F 96365 (inhibitor of SOC and nonselective cation channel type-2 (NSCC-2)), LOE 908 (inhibitor of NSCC-1 and NSCC-2), and La(3+) (inhibitor of transient receptor potential canonical (TRPC) channel). Reduction of extracellular Na(+) and pretreatment with KB-R7943, a Na(+)/Ca(2+) exchanger (NCX) inhibitor, inhibited both phases of [Ca(2+)](i) increases. These results suggest that 1) stimulation of alpha(1A)-AR with NA elicits the transient and sustained increases in [Ca(2+)](i) mediated through NSCC-2 that belongs to a TRPC family; 2) Na(+) influx through these channels drives NCX in the reverse mode, causing Ca(2+) influx in exchange for Na(+) efflux; and 3) the G(alphaq/11)/PLC/PKC-dependent pathway plays an important role in the increases in [Ca(2+)](i). [Abstract/Link to Full Text]

Utsu Y, Shinomiya K, Tokunaga S, Ohmori A, Kamei C
Effect of tandospirone on sleep latency in rats placed on a grid suspended over water.
J Pharmacol Sci. 2007 Sep;105(1):112-6.
The present study was performed to examine the effect of tandospirone on sleep latency in a new insomnia animal model by placing rats on a grid suspended over water. For investigating the mechanism of tandospirone, the effect of tandospirone on sleep latency was also studied using rats that were depleted with neuronal serotonin (5-HT) after p-chlorophenylalanine administration. Tandospirone caused a shortening of sleep latency dose-dependently, and a significant effect was observed at 20 mg/kg, p.o. or more. A shortening of sleep latency was observed by administration of p-chlorophenylalanine (300 mg/kg, i.p.) for 2 days. On the other hand, tandospirone exerted no potentiating effect on the shortening of sleep latency induced by p-chlorophenylalanine. From these findings, a shortening of sleep latency induced by tandospirone may occur through the pre-synaptic 5-HT(1A) receptors in rats. [Abstract/Link to Full Text]

Nyan M, Sato D, Oda M, Machida T, Kobayashi H, Nakamura T, Kasugai S
Bone formation with the combination of simvastatin and calcium sulfate in critical-sized rat calvarial defect.
J Pharmacol Sci. 2007 Aug;104(4):384-6.
Simvastatin, a cholesterol synthesis inhibitor, enhances BMP2 expression in osteoblasts. The purpose of the present study was to examine whether simvastatin stimulates bone regeneration when combined with calcium sulfate as a carrier. Critical-sized bone defects in rat calvaria were treated with calcium sulfate or with combination of 1 mg simvastatin and calcium sulfate. In the combination group, although the least amount of bone formation with intense soft tissue inflammation was observed at 2 and 4 weeks, remarkable bone formation was evident at 8 weeks. Conclusively, the combination of simvastatin and calcium sulfate stimulated bone regeneration in spite of the inflammatory response. [Abstract/Link to Full Text]

Ban JO, Yuk DY, Woo KS, Kim TM, Lee US, Jeong HS, Kim DJ, Chung YB, Hwang BY, Oh KW, Hong JT
Inhibition of cell growth and induction of apoptosis via inactivation of NF-kappaB by a sulfurcompound isolated from garlic in human colon cancer cells.
J Pharmacol Sci. 2007 Aug;104(4):374-83.
Compounds such as S-allylmercaptocysteine, diallyl disulfide, and S-trityl-L-cysteine isolated from garlic have been known to be effective in chemoprevention. Nuclear transcription factor-kappaB (NF-kappaB) has been known to be an implicated factor in apoptotic cell death of several cancer cells. In this study, we investigated whether a sulfurcompound (named thiacremonone) isolated from garlic could modulate NF-kappaB activity and thereby induce apoptotic cell death of colon cancer cells. Treatment with different concentrations (30 - 150 microg/ml) of thiacremonone for various periods (0 - 48 h) inhibited colon cancer cell (SW620 and HCT116) growth followed by induction of apoptosis in a dose-dependent manner. We also found that thiacremonone modulated tumor necrosis factor-alpha (TNF-alpha) and tetradeanoyl phorbol acetate (TPA)-induced NF-kappaB transcriptional and DNA binding activity. Moreover, thiacremonone suppressed NF-kappaB target anti-apoptotic genes (Bcl-2, cIAP1/2, and XIAP) and inflammatory genes (iNOS and COX-2), whereas it induced apoptotic genes (Bax, cleaved caspse-3, and cleaved PARP) expression. These results suggest that a novel sulfurocompound from garlic inhibited colon cancer cell growth through induction of apoptotic cell death by modulating of NF-kappaB. [Abstract/Link to Full Text]

Sukmawan R, Yada T, Toyota E, Neishi Y, Kume T, Shinozaki Y, Mori H, Ogasawara Y, Kajiya F, Yoshida K
Edaravone preserves coronary microvascular endothelial function after ischemia/reperfusion on the beating canine heart in vivo.
J Pharmacol Sci. 2007 Aug;104(4):341-8.
We examined whether edaravone (3-methyl-1-phenyl-2-pyrazolin-5-one), a free radical scavenger, exerts its protective effect on coronary microvessels after ischemia/reperfusion (I/R) in vivo. Ninety-minute coronary occlusion followed by reperfusion was performed in 16 open-chest dogs with and without edaravone administration. Coronary small artery (> or = 100 microm in size) and arteriolar (< 100 microm) vasodilation, in the presence of endothelium-dependent (acetylcholine) or -independent (papaverine) vasodilators, was directly observed using intravital microscopy before and after I/R. I/R impaired microvascular vasodilation in response to acetylcholine, whereas administration of edaravone preserved the response in microvessels of both sizes, but to a greater extent in the coronary small arteries. No significant changes were noted with papaverine administration. In the edaravone group, the fluorescent intensity from reactive oxygen species (ROS) was lower, whereas nitric oxide (NO) intensity was higher relative to controls in the microvessels of the ischemic area. In conclusion, edaravone preserves coronary microvascular endothelial function after I/R in vivo. These effects, which were NO-mediated, were attributed to the ROS scavenging properties of edaravone. [Abstract/Link to Full Text]

Takahashi-Yanaga F, Sasaguri T
The Wnt/beta-catenin signaling pathway as a target in drug discovery.
J Pharmacol Sci. 2007 Aug;104(4):293-302.
The cell signaling cascades provoked by Wnt proteins (the Wnt signaling pathways), which are well conserved through evolution, play crucial roles to maintain homeostasis of a variety of tissues such as skin, blood, intestine, and brain, as well as to regulate proliferation, morphology, motility, and fate of cells during embryonic development. Among these pathways, the signal transduction through beta-catenin (the Wnt/beta-catenin signaling pathway) has been most intensively studied because this signal regulates the expression of a number of genes essential for cell proliferation and differentiation and also this pathway is perturbed in a number of diseases such as cancers, bone diseases, and cardiovascular diseases. However, there is no therapeutic agents that can selectively modulate the Wnt/beta-catenin signaling pathway, although some existing drugs (e.g., non-steroidal anti-inflammatory drugs, vitamins, and imatinib mesylate) have been suggested to inhibit this pathway. Here we provide an overview of the Wnt/beta-catenin signaling pathway: its roles in physiology and pathology and the possibility as a target in development of new drugs. [Abstract/Link to Full Text]

Kanematsu T, Mizokami A, Watanabe K, Hirata M
Regulation of GABA(A)-receptor surface expression with special reference to the involvement of GABARAP (GABA(A) receptor-associated protein) and PRIP (phospholipase C-related, but catalytically inactive protein).
J Pharmacol Sci. 2007 Aug;104(4):285-92.
GABA(A) receptors are heteropentameric ligand-gated chloride channels composed of a variety of subunits, including alpha1 - 6, beta1 - 3, gamma1 - 3, delta, epsilon, theta, and pi, and play a key role in controlling inhibitory neuronal activity. Modification of the efficacy of the synaptic strength is produced by changes in both the number of neuronal surface receptors and pentameric molecular assembly, leading to differences of sensitivity to neurotransmitters and neuromimetic drugs. Therefore, it is important to understand the molecular mechanisms regulating the so-called "life cycle of GABA(A) receptors" including sequential pentameric assembly at the site synthesized, intracellular transport through the Golgi apparatus and the cytoplasm, insertion into the cell membrane, functional modulation at the cell surface, and finally internalization, followed by either recycling back to the surface membrane or lysosomal degradation. This review is focused on events related to the surface expression of the receptor containing the gamma2 subunit and clathrin/AP2 complex-mediated phospho-regulated endocytosis of the receptor, with special reference to the function of novel GABA(A) receptor modulators, GABARAP (GABA(A) receptor-associated protein) and PRIP (phospholipase C-related, but catalytically inactive protein). [Abstract/Link to Full Text]

Enomoto R, Sugahara C, Suzuki C, Nagase I, Takamura Y, Yoshikawa A, Hosoda A, Hirano H, Yokoi T, Lee E
Wogonin prevents glucocorticoid-induced thymocyte apoptosis without diminishing its anti-inflammatory action.
J Pharmacol Sci. 2007 Aug;104(4):355-65.
The effect of wogonin, a flavone highly purified from the roots of Scutellaria baicalensis, on apoptotic cell death was re-evaluated in rat thymocytes. This flavone inhibited glucocorticoid-induced apoptotic changes such as DNA fragmentation, phosphatidylserine translocation, and nuclear condensation in rat thymocytes. Similar inhibition was also observed in apoptosis induced by other inducers such as etoposide. No significant changes of these apoptotic features were observed in rat thymocytes treated with wogonin alone, suggesting that this flavone protects against glucocorticoid-mediated immunosuppression caused by thymocyte apoptosis. Wogonin was reported to possess anti-inflammatory action in some previous studies, but this flavone had no effect on carrageenan-induced paw edema in this study. The simultaneous treatment of wogonin and glucocorticoid neither enhanced nor reduced the anti-inflammatory effect of glucocorticoid. These results indicate that wogonin is likely to prevent the immunosuppression of glucocorticoid without diminishing its drug efficacy as an anti-inflammatory agent. [Abstract/Link to Full Text]

Iida-Tanaka N, Namekata I, Kaneko M, Tamura M, Kawanishi T, Nakamura R, Shigenobu K, Tanaka H
Involvement of intracellular Ca(2+) in the regulatory volume decrease after hyposmotic swelling in MDCK cells.
J Pharmacol Sci. 2007 Aug;104(4):397-401.
We examined the source of Ca(2+) involved in the volume regulation of Madin-Darby canine kidney (MDCK) cells with confocal microscopy and fluoroprobes. Hyposmosis induced a transient increase in cell volume, as well as cytoplasmic Ca(2+), which peaked at 3 to 5 min and gradually decreased to reach the initial value within about 30 min. This late decrease in cell volume, as well as the transient rise in cytoplasmic Ca(2+), was reduced in Ca(2+)-free solution and was abolished by pretreatment with thapsigargin. In conclusion, Ca(2+) released from the intracellular store contributes to the regulatory volume decrease following hyposmotic swelling in MDCK cells. [Abstract/Link to Full Text]

Oda T, Kume T, Katsuki H, Niidome T, Sugimoto H, Akaike A
Donepezil potentiates nerve growth factor-induced neurite outgrowth in PC12 cells.
J Pharmacol Sci. 2007 Aug;104(4):349-54.
Donepezil is a potent and selective acetylcholinesterase inhibitor developed for the treatment of Alzheimer's disease. To elucidate whether donepezil causes neuronal differentiation, we examined its effect on nerve growth factor (NGF)-induced neurite outgrowth in PC12 cells. Donepezil (10 microM) significantly potentiated the neurite outgrowth evoked by low (1 ng/ml) and high (50 ng/ml) concentrations of NGF. The effect of donepezil (1 - 10 microM) was concentration-dependent. The enhancement of neurite outgrowth caused by donepezil was not blocked by the acetylcholine receptor (AChR) antagonists mecamylamine and scopolamine. Furthermore, the AChR agonists nicotine and carbachol did not affect the neurite outgrowth induced by NGF. Donepezil (10 microM) also significantly potentiated neurite outgrowth evoked by dibutyryl cyclic AMP. Moreover, donepezil potentiated the NGF-induced phosphorylation of extracellular signal-regulated kinase (ERK). These results suggest that donepezil potentiated neuronal differentiation by enhancing the activation of ERK. [Abstract/Link to Full Text]

Ishii T, Ishimori H, Mori K, Uto T, Fukuda K, Urashima T, Nishimura M
Bovine lactoferrin stimulates anchorage-independent cell growth via membrane-associated chondroitin sulfate and heparan sulfate proteoglycans in PC12 cells.
J Pharmacol Sci. 2007 Aug;104(4):366-73.
Bovine lactoferrin (bLf) is an iron-binding secretory protein present in breast milk, mucosal secretions, and the secondary granules of neutrophils. Although bLf has multiple functions, including antimicrobial and immunomodulatory activities, its effect on neuronal cells is not fully understood. We report that bLf prevents cell adhesion of PC12 cells and allows them to be cultivated in suspension. PC12 cells normally adhere well to plastic culture plates and show anchorage-dependent cell growth, but we found that soon after adding bLf, they detach from culture plates and begin to grow in suspension. When bLf was removed from the medium, the cells began to re-adhere to the plates. Thus, bLf inhibits cell adhesion and stimulates anchorage-independent growth in PC12 cells. On the other hand, bLf-induced cell suspension growth was not observed when cells were grown on a laminin matrix, suggesting that bLf does not affect integrin-mediated cell adhesion on a laminin matrix. Treatment of cells with heparin or chondroitin sulfate A or C inhibited bLf-induced growth in cell suspension. Furthermore, pretreatment of cells with heparinase and/or chondroitinase prevented direct binding of bLf to the cell membrane. These results suggest that bLf binds to the membrane of PC12 cells via membrane-associated proteoglycans and leads to anchorage-independent growth. [Abstract/Link to Full Text]

Ichikawa J, Yamada RX, Muramatsu R, Ikegaya Y, Matsuki N, Koyama R
Cryopreservation of granule cells from the postnatal rat hippocampus.
J Pharmacol Sci. 2007 Aug;104(4):387-91.
Although primary cultures of neurons are essential methods for cell biological and pharmacological researches, many animals must be sacrificed for each experiment. Here we introduce a novel system to cryopreserve hippocampal granule cells (GCs) prepared from postnatal rats. Being thawed after as long as 60 days of cryopreservation, GCs expressed the mature neuronal marker MAP-2 and elongated single tau-1-positive axons and multiple tau-1-negative dendrites. These properties closely resembled intact GCs in primary cultures, providing the advantage of being able to repeatedly prepare stable cultures with a single sacrifice of animals. [Abstract/Link to Full Text]

Kubo S, Doe I, Kurokawa Y, Kawabata A
Hydrogen sulfide causes relaxation in mouse bronchial smooth muscle.
J Pharmacol Sci. 2007 Aug;104(4):392-6.
We investigated the effects of NaHS, a hydrogen sulfide (H(2)S) donor, on the tension of isolated mouse and guinea-pig bronchial rings. NaHS at 0.01 - 10 mM had no effect on the tone of those preparations without precontraction. When the preparation was precontracted with carbachol, NaHS at 0.1 - 3 mM strongly relaxed the mouse rings, but produced only slight relaxation in the guinea-pig rings. The NaHS-induced relaxation in the mouse bronchus was resistant to inhibitors of ATP-sensitive K(+) channels, soluble guanylyl cyclase, cyclooxygenase (COX)-1 or COX-2, and antagonists of tachykinin receptors. Thus, NaHS evokes bronchodilation in mice, suggesting a possible role for H(2)S in the respiratory system. [Abstract/Link to Full Text]

Hirabara Y, Araki M, Fukuda M, Katafuchi S, Honda K, Saito R, Takano Y
A high-sodium diet in streptozotocin-induced diabetic rats impairs endothelium-derived hyperpolarizing factor-mediated vasodilation.
J Pharmacol Sci. 2007 Aug;104(4):402-5.
We examined endothelium-derived hyperpolarizing factor (EDHF)-mediated relaxation of mesenteric arteries in high-sodium loaded streptozotocin (STZ)-induced diabetic rats. The study shows that acetylcholine (ACh)-induced, EDHF-mediated relaxation is relatively maintained in STZ-induced diabetic rats, but after a high-sodium diet was given, the function was significantly impaired in STZ-induced diabetic rats. [Abstract/Link to Full Text]

Tokita Y, Yuzurihara M, Sakaguchi M, Satoh K, Kase Y
The pharmacological effects of Daikenchuto, a traditional herbal medicine, on delayed gastrointestinal transit in rat postoperative ileus.
J Pharmacol Sci. 2007 Aug;104(4):303-10.
The effect of Daikenchuto, a traditional herbal medicine, on gastrointestinal hypoperistalsis in postoperative ileus (POI) was investigated. POI was induced by laparotomy with manipulation of the gastrointestine under anesthesia, and gastrointestinal transit was calculated by migration of Evans blue. Daikenchuto (270 - 2,700 mg/kg, p.o.) dose-dependently improved the delayed gastrointestinal transit in POI. This effect of Daikenchuto was partially inhibited by SB204070 (1 mg/kg, s.c.), a 5-hydroxytriptamine(4) (5-HT(4))-receptor antagonist and completely abolished by atropine (1 mg/kg, s.c.), a muscarine-receptor antagonist. Among the constituents of Daikenchuto, the medical herb zanthoxylum fruit (60 mg/kg, p.o.) and maltose syrup (2,400 mg/kg, p.o.) significantly ameliorated the delayed gastrointestinal transit, but ginseng and processed ginger did not affect the gastrointestinal transit in the rat POI. The improvement induced by zanthoxylum fruit was also inhibited by atropine or SB204070. In addition, the high osmotic pressure of the maltose syrup (2400 mg/10 mL per kg) was related to the improvement of delayed gastrointestinal transit. These results demonstrated that Daikenchuto ameliorates postoperative hypoperistalsis via cholinergic nerves and 5-HT(4) receptors and that osmotic pressure also may be involved in this action. Moreover, zanthoxylum fruit and maltose syrup were crucial medical herbs contributing to the ability of Daikenchuto. [Abstract/Link to Full Text]

Kaneda T, Konno Y, Urakawa N, Nakajyo S, Shimizu K
Ibudilast-induced decreases in cytosolic Ca(2+) level and contraction in rat aorta.
J Pharmacol Sci. 2007 Aug;104(4):311-8.
The mechanism by which ibudilast induces vasodilation was examined in isolated endothelium-denuded rat aorta. Ibudilast inhibited the contractions induced by phenylephrine (PE) and high K(+) with decrease of [Ca(2+)](i) level in a concentration-dependent manner, to the same degree. 3-Isobutyl-1-methylxanthine (IBMX) inhibited PE-induced contraction and [Ca(2+)](i) level in a concentration-dependent manner, but it inhibited high K(+)-induced contraction without decrease of [Ca(2+)](i) level. In comparison with IBMX, the increases of cAMP and cGMP contents in ibudilast were much smaller than that of muscle tension. Ibudilast did not inhibit 12-deoxyphorbol 13-isobutyrate (DPB)-induced contraction in the presence of verapamil. Treatment with 30 microM ibudilast inhibited the extracellularly added Ca(2+)-induced muscle tension and increases in [Ca(2+)](i) level during high K(+) depolarization. These results suggested that ibudilast inhibited PE- and high K(+)-induced muscle contractions mainly by the inhibition of [Ca(2+)](i) level in endothelium-denuded rat aorta. [Abstract/Link to Full Text]

Takei H, Iizuka S, Yamamoto M, Takeda S, Yamamoto M, Arishima K
The herbal medicine Tokishakuyakusan increases fetal blood glucose concentrations and growth hormone levels and improves intrauterine growth retardation induced by N(omega)-nitro-L-arginine methyl ester.
J Pharmacol Sci. 2007 Aug;104(4):319-28.
N(omega)-Nitro-L-arginine methyl ester (L-NAME) induces a pre-eclampsia-like syndrome in pregnant rats. We have previously reported the anti-hypertensive effects of several Japanese traditional (Kampo) medicines in this model, and one of these, Tokishakuyakusan (TS), also improved intrauterine growth retardation (IUGR). In the present study, we characterized the effect of TS on IUGR. TS administration reversed the decrease in fetal body weight and fetal blood glucose concentration induced by the infusion of L-NAME. Growth hormone (GH) levels in the fetal blood, which were decreased by L-NAME infusion, were also significantly elevated by TS; however, levels of GH releasing hormone (GHRH) and insulin-like growth factor I (IGF-I) were unchanged and only slightly changed, respectively. Treatment with L-NAME with or without TS had no apparent effect on GH, GHRH, and IGF-I levels of dams. In an immunocytochemical study, the number of GH-positive cells in the fetal pituitary gland was significantly increased in TS-treated rats. These data suggest that enhanced proliferation of somatotrope cells of the pituitary gland and the resultant increase in GH secretion in the fetus may be involved in the improvement of IUGR by TS. [Abstract/Link to Full Text]

Pu F, Mishima K, Irie K, Motohashi K, Tanaka Y, Orito K, Egawa T, Kitamura Y, Egashira N, Iwasaki K, Fujiwara M
Neuroprotective effects of quercetin and rutin on spatial memory impairment in an 8-arm radial maze task and neuronal death induced by repeated cerebral ischemia in rats.
J Pharmacol Sci. 2007 Aug;104(4):329-34.
In order to determine the differential effects of flavonoids on cerebral ischemia, we investigated the effects of (-)-epigallocatechin gallate (EGCG), catechin, rutin, and quercetin on spatial memory impairment and neuronal death induced by repeated cerebral ischemia in rats. Both rutin and quercetin (50 mg/kg x 2) improved spatial memory impairment in the 8-arm radial maze task and neuronal death in the hippocampal CA1 area; however, catechin (200 mg/kg x 2) and EGCG (50 mg/kg x 1) did not. Administration of EGCG (50 mg/kg x 2) resulted in a high mortality rate. These results suggest that in this repeated cerebral ischemia model, the 4-oxo group and the 2,3-double bond in the C ring of rutin and quercetin are related to their neuroprotective action. [Abstract/Link to Full Text]

Recent Articles in Alternative Medicine Review

Boger RH
Letter to the editor re: JAMA article on L-arginine therapy in acute myocardial infarction.
Altern Med Rev. 2006 Jun;11(2):91-2. [Abstract/Link to Full Text]

Head K, Miller A
Bad medicine, bad reportage, or both?
Altern Med Rev. 2006 Jun;11(2):83-90. [Abstract/Link to Full Text]

Molybdenum. Monograph.
Altern Med Rev. 2006 Jun;11(2):156-61. [Abstract/Link to Full Text]

Monograph. Eleutherococcus senticosus.
Altern Med Rev. 2006 Jun;11(2):151-5. [Abstract/Link to Full Text]

Spelman K, Burns J, Nichols D, Winters N, Ottersberg S, Tenborg M
Modulation of cytokine expression by traditional medicines: a review of herbal immunomodulators.
Altern Med Rev. 2006 Jun;11(2):128-50.
Modulation of cytokine secretion may offer novel approaches in the treatment of a variety of diseases. One strategy in the modulation of cytokine expression may be through the use of herbal medicines. A class of herbal medicines, known as immunomodulators, alters the activity of immune function through the dynamic regulation of informational molecules such as cytokines. This may offer an explanation of the effects of herbs on the immune system and other tissues. For this informal review, the authors surveyed the primary literature on medicinal plants and their effects on cytokine expression, taking special care to analyze research that utilized the multi-component extracts equivalent to or similar to what are used in traditional medicine, clinical phytotherapy, or in the marketplace. METHODOLOGY: MEDLINE, EBSCO, and BIOSIS were used to identify research on botanical medicines, in whole or standardized form, that act on cytokine activity through different models, i.e., in vivo (human and animal), ex vivo, or in vitro. RESULTS: Many medicinal plant extracts had effects on at least one cytokine. The most frequently studied cytokines were IL-1, IL-6, TNF, and IFN. Acalypha wilkesiana, Acanthopanax gracilistylus, Allium sativum, Ananus comosus, Cissampelos sympodialis, Coriolus versicolor, Curcuma longa, Echinacea purpurea, Grifola frondosa, Harpagophytum procumbens, Panax ginseng, Polygala tenuifolia, Poria cocos, Silybum marianum, Smilax glabra, Tinospora cordifolia, Uncaria tomentosa, and Withania somnifera demonstrate modulation of multiple cytokines. CONCLUSION: The in vitro and in vivo research demonstrates that the reviewed botanical medicines modulate the secretion of multiple cytokines. The reported therapeutic success of these plants by traditional cultures and modern clinicians may be partially due to their effects on cytokines. Phytotherapy offers a potential therapeutic modality for the treatment of many differing conditions involving cytokines. Given the activity demonstrated by many of the reviewed herbal medicines and the increasing awareness of the broad-spectrum effects of cytokines on autoimmune conditions and chronic degenerative processes, further study of phytotherapy for cytokine-related diseases and syndromes is warranted. [Abstract/Link to Full Text]

Patrick L
Lead toxicity part II: the role of free radical damage and the use of antioxidants in the pathology and treatment of lead toxicity.
Altern Med Rev. 2006 Jun;11(2):114-27.
Lead is an environmentally persistent toxin that causes neurological, hematological, gastrointestinal, reproductive, circulatory, and immunological pathologies. The propensity for lead to catalyze oxidative reactions and generate reactive oxygen species has been demonstrated in multiple studies. These reactive oxygen species (ROS) inhibit the production of sulfhydryl antioxidants, inhibit enzyme reactions impairing heme production, cause inflammation in vascular endothelial cells, damage nucleic acids and inhibit DNA repair, and initiate lipid peroxidation in cellular membranes. These wide-ranging effects of ROS generation have been postulated to be major contributors to lead-exposure related disease. Antioxidants - vitamins B6, C and E, zinc, taurine, N-acetylcysteine, and alpha-lipoic acid, either alone or in conjunction with standard pharmaceutical chelating agents - have been studied in lead-exposed animals. The evidence for their use in lead exposure, alone and in conjunction with chelating agents, is reviewed in this article. [Abstract/Link to Full Text]

Roxas M
Herpes zoster and postherpetic neuralgia: diagnosis and therapeutic considerations.
Altern Med Rev. 2006 Jun;11(2):102-13.
Herpes zoster (HZ), also known as shingles, is a painful vesicular rash resulting from reactivation of the virus that also causes chickenpox - Varicella zoster virus (VZV). Typically, the rash runs its course in a matter of 4-5 weeks. The pain, however, may persist months, even years, after the skin heals. This phenomenon is known as postherpetic neuralgia (PHN). Often described as an intense burning, itching sensation, this pain can be significant to the point of being debilitating, and as such can greatly affect quality of life. Although shingles is generally regarded as a self-limited condition, the fact it can take several weeks to resolve and has the potential for development of complications such as PHN presents a challenge to clinicians. Many treatment options are available, each offering variable levels of efficacy. Conventional therapies include prescription antivirals, corticosteroids, and analgesics, both oral and topical. Other considerations include use of over-the-counter anti-inflammatory agents, physiotherapy, and nerve block injections. This article reviews herpes zoster and postherpetic neuralgia, and presents the most effective conventional treatment options currently available, as well as select botanical, nutritional, and other considerations that may be beneficial in the management of this condition. [Abstract/Link to Full Text]

Gaby AR
Natural remedies for Herpes simplex.
Altern Med Rev. 2006 Jun;11(2):93-101.
Herpes simplex is a common viral infection of the skin or mucous membranes. The lesions caused by this infection are often painful, burning, or pruritic, and tend to recur in most patients. Short-term treatment with acyclovir can accelerate the healing of an acute outbreak, and continuous acyclovir therapy is often prescribed for people with frequent recurrences. While this drug can reduce the recurrence rate by 60-90 percent, it can also cause a wide array of side effects, including renal failure, hepatitis, and anaphylaxis. Safe and effective alternatives are therefore needed. There is evidence that certain dietary modifications and natural substances may be useful for treating active Herpes simplex lesions or preventing recurrences. Treatments discussed include lysine, vitamin C, zinc, vitamin E, adenosine monophosphate, and lemon balm (Melissa officinalis). [Abstract/Link to Full Text]

L-Tryptophan. Monograph.
Altern Med Rev. 2006 Mar;11(1):52-6. [Abstract/Link to Full Text]

Coleus forskohlii. Monograph.
Altern Med Rev. 2006 Mar;11(1):47-51. [Abstract/Link to Full Text]

Friel PN, Alexander T, Wright JV
Suppression of adrenal function by low-dose prednisone: assessment with 24-hour urinary steroid hormone profiles--a review of five cases.
Altern Med Rev. 2006 Mar;11(1):40-6.
The impact of the synthetic glucocorticoid prednisone on adrenal steroid hormone production was examined using 24-hour urinary steroid hormone profiling. Five women, who were chronically taking low-dose prednisone, were tested, and the relevant literature was reviewed. As expected, adrenal glucocorticoid production, measured by urinary terminal cortisol and cortisone metabolites, was markedly suppressed compared to reference range values (p=0.03). Urinary cortisol and cortisone, reflecting circulating glucocorticoids, were decreased to a lesser extent than their terminal metabolites. Urinary dehydroepiandrosterone (DHEA) excretion was dramatically suppressed (p=0.03), while the downstream androgen metabolites androsterone and etiocholanolone were suppressed to a lesser extent. Aldosterone and tetrahydrocorticosterone production demonstrated modest suppression after prednisone administration, but allo-tetrahydrocorticosterone, which is highly sensitive to adrenocorticotropic hormone (ACTH) secretion, was suppressed to a greater extent. Prednisone administration results in a decrease in ACTH secretion by the anterior pituitary, suppressing synthesis of glucocorticoids, DHEA, and DHEA metabolites. Decreased glucocorticoid synthesis is adaptive, because prednisone is active at the glucocorticoid receptor, but suppression of DHEA synthesis is not mitigated by prednisone. DHEA is an important sex hormone precursor, neurosteroid, and endocrine and immune modulator; therefore, DHEA depletion may have significant adverse consequences in terms of sex hormone production, bone health, endocrine and immune system function, and neuropsychiatric status. Studies of DHEA replacement in patients taking prednisone for lupus demonstrate amelioration of some of these adverse effects. [Abstract/Link to Full Text]

Bradley R, Oberg EB
Naturopathic medicine and type 2 diabetes: a retrospective analysis from an academic clinic.
Altern Med Rev. 2006 Mar;11(1):30-9.
Accurate descriptions of naturopathic medicine as a whole system of medical practice are rare in the literature and non-existent for type 2 diabetes. METHODS: Using retrospective analysis of medical records at an academic naturopathic outpatient clinic, data was abstracted to investigate patterns of patient status, details of treatment recommendations, and levels of evidence. RESULTS: Most naturopathic medical care for type 2 diabetes is adjunctive, although naturopathic physicians are qualified to fill the role of primary care providers. Glycemic control and other vital statistics in patients receiving naturopathic care are comparable to published national averages. Naturopathic physicians prescribe comprehensive therapeutic lifestyle change recommendations supported by a high level of evidence - 100 percent received dietary counseling, 69 percent were taught stress reduction techniques, and 94 percent were prescribed exercise. Patients additionally received prescriptions for botanical and nutritional supplementation, often in combination with conventional medication. Analysis of individual supplement effects was not performed due to inadequate records. Components of treatment recommendations are often evidence-based, with varying evidence quality. CONCLUSION: Naturopathic medicine as a whole medical system supplies evidence-based lifestyle recommendations as suggested in management guidelines for diabetes, hypertension, and hyperlipidemia set forth by the respective national organizations - the American Diabetes Association (ADA), the Joint National Committee on Hypertension (JNC-7), and the National Cholesterol Education Program results of the third Adult Treatment Panel (NCEP ATP-III). Increased research effort to determine the safety and efficacy of combinations of supplements or medications and supplements is warranted. Education of other health care providers, patients, and health policy makers regarding the value of the naturopathic approach in the treatment and prevention of type 2 diabetes is warranted, yet prospective data on efficacy must be collected. [Abstract/Link to Full Text]

Miller AL
The effects of sustained-release-L-arginine formulation on blood pressure and vascular compliance in 29 healthy individuals.
Altern Med Rev. 2006 Mar;11(1):23-9.
Vascular endothelial function is crucial to cardiovascular function and thus to blood perfusion to the heart and throughout the body. A number of substances are produced and secreted by vascular endothelial cells, the most important of which is nitric oxide, a potent regulator of vascular function. Nitric oxide diffuses from endothelial cells into underlying smooth muscle, causing relaxation, which results in vasodilation. When this process is inhibited or inadequate the arteries cannot dilate as necessary, resulting in hypertonicity and reduced blood flow. Such endothelial dysfunction also causes increased platelet and monocyte adhesiveness and smooth muscle proliferation, processes thought to be at the genesis of atherosclerotic plaque formation. Since L-arginine is the body's only substrate for nitric oxide synthesis, adequate L-arginine must be present for proper nitric oxide production. In this open label trial, a group of 29 asymptomatic individuals were given L-arginine (1,050 mg, as Perfusia-SR, a sustained-release preparation) twice daily (total 2.1 g daily) for one week. Systolic blood pressure was reduced in 62 percent of participants compared to baseline, with a non-significant mean decrease in all patients of 4 mm Hg. Diastolic blood pressure was reduced in 69 percent of participants, with a mean reduction of 3.7 mm Hg (p = 0.005). In the 10 individuals who were borderline or hypertensive (systolic > 130 or diastolic > 85), there was a mean systolic reduction of 11 mm Hg (p = 0.05), while normotensives (n = 19) had a mean systolic decrease of only 0.22 mm Hg. Diastolic blood pressure was decreased a non-significant 4.9 mm Hg in borderline or hypertensives and 4.5 mm Hg in normotensives (p = 0.026). Vascular elasticity relates to endothelial function, and can be measured non-invasively. At baseline and follow-up, vascular compliance was assessed via digital pulse wave analysis (DPA; Meridian Medical). After one week, pulse wave analysis showed a significant increase in large artery compliance (mean 23% improvement; p = 0.02) and a non-significant increase in small artery compliance (mean 23% improvement; p = 0.15). This study demonstrates blood pressure reductions, especially in patients with borderline or frank hypertension, as well as improved vascular compliance - an indicator of improved endothelial function and perfusion - after a one-week trial of sustained-release L-arginine. Poor endothelial function due to inadequate endothelial nitric oxide production is present in hypertension, as well as in numerous other aspects of cardiovascular disease, including angina, erectile dysfunction, cerebrovascular disease, and peripheral vascular disease. This is the first study showing a moderate dose of sustained-release L-arginine can improve endothelial function and blood pressure. [Abstract/Link to Full Text]

Patrick L
Lead toxicity, a review of the literature. Part 1: Exposure, evaluation, and treatment.
Altern Med Rev. 2006 Mar;11(1):2-22.
The phasing out of leaded gasoline for transportation vehicles between 1973 and 1995 and the removal of lead from paint by federal mandate by 1978 have resulted in substantial lowering of mean blood lead levels in all segments of the U.S. population. However, because lead is a persistent metal, it is still present in the environment - in water, brass plumbing fixtures, soil, dust, and imported products manufactured with lead. Diagnosis of lead toxicity has traditionally been based on significantly elevated blood lead levels. However, data now implicates low-level exposures and blood lead levels previously considered normal as causative factors in cognitive dysfunction, neurobehavioral disorders, neurological damage, hypertension, and renal impairment. Chelation is the conventional recommendation in the case of blood levels associated with acute toxicity and encephalopathic damage. Issues surrounding the assessment of body lead burden and the consequences of low-level environmental exposure are critical in the treatment of chronic disease related to lead toxicity. [Abstract/Link to Full Text]

Czp A
Citrulline, Viagra and BiDil--bad medicine.
Altern Med Rev. 2005 Dec;10(4):265-7. [Abstract/Link to Full Text]

Indole-3-carbinol. Monograph.
Altern Med Rev. 2005 Dec;10(4):337-42. [Abstract/Link to Full Text]

Melatonin. Monograph.
Altern Med Rev. 2005 Dec;10(4):326-36. [Abstract/Link to Full Text]

Roberts AT, de Jonge-Levitan L, Parker CC, Greenway F
The effect of an herbal supplement containing black tea and caffeine on metabolic parameters in humans.
Altern Med Rev. 2005 Dec;10(4):321-5.
OBJECTIVE: The objective of this study was to test an herbal supplement containing black tea (the fully oxidized form of Camellia sinensis) and caffeine for stimulation of thermogenesis. METHODS/MATERIALS: A double-blind, placebo-controlled, crossover study was conducted on 16 healthy, weight-stable, non-smoking subjects, ages 21-55 years, with body mass index (BMI) of 20-30 kg/m2, and on no medications other than oral contraceptives or hormone replacement therapy. Subjects had no caffeine for 48 hours, no exercise for 24 hours, and no food for 12 hours before each visit. Area under the curve (AUC) for resting metabolic rate (RMR), respiratory quotient (RQ), blood pressure, pulse rate, and temperature were measured. At each visit RMR was measured at baseline and at one and two hours following oral administration of a supplement containing principally 600 mg black tea extract (60 percent polyphenols, 20 percent caffeine) and 442 mg guarana extract (36 percent caffeine) or matching placebo. RESULTS: The RMR and systolic blood pressure (SBP) AUCs increased significantly (p less than 0.02 and p less than 0.01, respectively) in the herbal supplement group compared to placebo. The AUC increase in RMR over the two-hour test period was 77.19 kcal/24 hr2 +/- 120.10 kcal/24 hr2 with an average rise of 52.38 +/- 29.52 kcal/24 hrs. The AUC rise in SBP over two hours was 10.3 mm Hg/hr +/- 14 mm Hg/hr. The average rise in SBP over two hours was 3.7 mm Hg +/- 4.4 mm Hg. DISCUSSION: The herbal supplement increased metabolic rate without changing substrate oxidation. The rise in SBP was consistent with the amount of caffeine the supplement contained. [Abstract/Link to Full Text]

Kelly GS
Seasonal variations of selected cardiovascular risk factors.
Altern Med Rev. 2005 Dec;10(4):307-20.
This article reviews research on selected biomarkers of cardiovascular risk - cholesterol and other lipids, C-reactive protein (CRP), fibrinogen, homocysteine - in the attempt to determine the existence of a predictable seasonal chronobiological pattern of variation. Studies dating as far back as the 1930s have reported seasonal variations in cholesterol levels. Statistically significant seasonal changes in lipid levels have been found in individuals irrespective of the country where the research has been conducted, and irrespective of the age, sex, ethnicity, and baseline lipid levels of the study subjects. While not all studies have been in complete agreement on either the amplitude (degree of seasonal change) or month/s of highest lipid levels, a strong winter/summer difference has been found in most studies. Existing evidence for an independent effect of season in variation of CRP is weak. Studies have consistently reported significant seasonal variations in fibrinogen levels. While other biological factors clearly interact to affect fibrinogen variability, seasonality appears to be an independent source of variability. Evidence from several studies points to a lack of seasonal variability in homocysteine levels. Although seasonal variability is just one source of periodicity influencing biological function and assessments in clinical practice, for some biomarkers, including lipids and fibrinogen, it is a source of variability that warrants consideration prior to a decision to treat and in assessing response to interventions. [Abstract/Link to Full Text]

Gaby AR
Adverse effects of dietary fructose.
Altern Med Rev. 2005 Dec;10(4):294-306.
The consumption of fructose, primarily from high-fructose corn syrup (HFCS), has increased considerably in the United States during the past several decades. Intake of HFCS may now exceed that of the other major caloric sweetener, sucrose. Some nutritionists believe fructose is a safer form of sugar than sucrose, particularly for people with diabetes mellitus, because it does not adversely affect blood-glucose regulation, at least in the short-term. However, fructose has potentially harmful effects on other aspects of metabolism. In particular, fructose is a potent reducing sugar that promotes the formation of toxic advanced glycation end-products, which appear to play a role in the aging process; in the pathogenesis of the vascular, renal, and ocular complications of diabetes; and in the development of atherosclerosis. Fructose has also been implicated as the main cause of symptoms in some patients with chronic diarrhea or other functional bowel disturbances. In addition, excessive fructose consumption may be responsible in part for the increasing prevalence of obesity, diabetes mellitus, and non-alcoholic fatty liver disease. Although the long-term effects of fructose consumption have not been adequately studied in humans, the available evidence suggests it may be more harmful than is generally recognized. The extent to which a person might be adversely affected by dietary fructose depends both on the amount consumed and on individual tolerance. With a few exceptions, the relatively small amounts of fructose that occur naturally in fruits and vegetables are unlikely to have deleterious effects, and this review is not meant to discourage the consumption of these healthful foods. [Abstract/Link to Full Text]

Kidd PM
Neurodegeneration from mitochondrial insufficiency: nutrients, stem cells, growth factors, and prospects for brain rebuilding using integrative management.
Altern Med Rev. 2005 Dec;10(4):268-93.
Degenerative brain disorders (neurodegeneration) can be frustrating for both conventional and alternative practitioners. A more comprehensive, integrative approach is urgently needed. One emerging focus for intervention is brain energetics. Specifically, mitochondrial insufficiency contributes to the etiopathology of many such disorders. Electron leakages inherent to mitochondrial energetics generate reactive oxygen free radical species that may place the ultimate limit on lifespan. Exogenous toxins, such as mercury and other environmental contaminants, exacerbate mitochondrial electron leakage, hastening their demise and that of their host cells. Studies of the brain in Alzheimer's and other dementias, Down syndrome, stroke, Parkinson's disease, multiple sclerosis, amyotrophic lateral sclerosis, Huntington's disease, Friedreich's ataxia, aging, and constitutive disorders demonstrate impairments of the mitochondrial citric acid cycle and oxidative phosphorylation (OXPHOS) enzymes. Imaging or metabolic assays frequently reveal energetic insufficiency and depleted energy reserve in brain tissue in situ. Orthomolecular nutrients involved in mitochondrial metabolism provide clinical benefit. Among these are the essential minerals and the B vitamin group; vitamins E and K; and the antioxidant and energetic cofactors alpha-lipoic acid (ALA), ubiquinone (coenzyme Q10; CoQ10), and nicotinamide adenine dinucleotide, reduced (NADH). Recent advances in the area of stem cells and growth factors encourage optimism regarding brain regeneration. The trophic nutrients acetyl L-carnitine (ALCAR), glycerophosphocholine (GPC), and phosphatidylserine (PS) provide mitochondrial support and conserve growth factor receptors; all three improved cognition in double-blind trials. The omega-3 fatty acid docosahexaenoic acid (DHA) is enzymatically combined with GPC and PS to form membrane phospholipids for nerve cell expansion. Practical recommendations are presented for integrating these safe and well-tolerated orthomolecular nutrients into a comprehensive dietary supplementation program for brain vitality and productive lifespan. [Abstract/Link to Full Text]

Czap AF
The Delilah revelation.
Altern Med Rev. 2005 Sep;10(3):169-71. [Abstract/Link to Full Text]

Glycyrrhiza glabra. Monograph.
Altern Med Rev. 2005 Sep;10(3):230-7. [Abstract/Link to Full Text]

Folic acid. Monograph.
Altern Med Rev. 2005 Sep;10(3):222-9. [Abstract/Link to Full Text]

Curcio JJ, Kim LS, Wollner D, Pockaj BA
The potential of 5-hydryoxytryptophan for hot flash reduction: a hypothesis.
Altern Med Rev. 2005 Sep;10(3):216-21.
Hormone replacement therapy (HRT) is contraindicated in women with a history of breast cancer or a high risk of breast cancer development. Recent results from large clinical trials, such as the Women's Health Initiative, have demonstrated increased risks of thromboembolic events and a moderate increased risk of breast cancer in women using conjugated estrogens and progestogens. There is a need for viable non-hormonal alternative treatments to HRT, such as nutritional and botanical therapies, in this population of women, who tend to experience more significant vasomotor symptoms. Safe and effective therapies that do not stimulate breast cell proliferation could prove extremely useful for the management of such symptoms for women in both low- and high-risk breast cancer populations. As a non-hormonal treatment, anti-depressants, such as selective serotonin reuptake inhibitors (SSRIs), have been shown to improve hot flash symptoms in women. The proposed mechanism is related to an increase in serotonin allowing for an increase in the set point of the brain's thermoregulator. In small clinical studies, the administration of tryptophan and 5-hydroxytryptophan (5HTP), the precursors of serotonin, have been shown to reduce depressive symptoms, possibly by enhancing the synthesis of serotonin. Thus, increased serotonin levels may have the ability to decrease hot flashes in a mechanism similar to that of SSRIs without the risks of breast cell stimulation. This would be particularly desirable for menopausal women with breast cancer or with risks of breast cancer. This article discusses the background information on hot flashes, SSRIs, tryptophan, and 5HTP, and possible clinical application of 5HTP for menopausal women with breast cancer risk. [Abstract/Link to Full Text]

Thomas R, Wilson D
Randomized controlled trials of non-medical and non-surgical therapies for palliative care: a literature review.
Altern Med Rev. 2005 Sep;10(3):204-15.
A systematic review of randomized controlled trials (RCTs) of non-medical and non-surgical therapies for palliative care was undertaken to provide guidance for best practice palliative care. Nine databases were searched (ERIC, EMBASE, MEDLINE, CINAHL, AHMED, Psychinfo, HealthStar, Sociological Abstracts, and the Cochrane Library, including Central and Systematic Reviews) for RCTs and systematic reviews. Fifteen RCTs, varied in intervention and outcome measures, were identified. Several studies found positive results but the sample sizes were small, the methodological quality of the RCTs did not meet Cochrane Collaboration criteria, and the conclusions were at high risk of bias. Improved planning of the protocols and execution, with the addition of experienced trialists and statisticians, is required to improve the quality of the evidence collected in future studies. [Abstract/Link to Full Text]

Kidd P, Head K
A review of the bioavailability and clinical efficacy of milk thistle phytosome: a silybin-phosphatidylcholine complex (Siliphos).
Altern Med Rev. 2005 Sep;10(3):193-203.
Certain of the water-soluble flavonoid molecules can be converted into lipid-compatible molecular complexes, aptly called phytosomes. Phytosomes are better able to transition from a hydrophilic environment into the lipid-friendly environment of the outer cell membrane, and from there into the cell, finally reaching the blood. The fruit of the milk thistle plant (Silybum marianum, Family Asteraceae) contains flavonoids that are proven liver protectants. The standardized extract known as silymarin contains three flavonoids of the flavonol subclass. Silybin predominates, followed by silydianin and silychristin. Although silybin is the most potent of the flavonoids in milk thistle, similar to other flavonoids it is not well-absorbed. Silybin-phosphatidylcholine complexed as a phytosome provides significant liver protection and enhanced bioavailability over conventional silymarin. [Abstract/Link to Full Text]

Helms S
Celiac disease and gluten-associated diseases.
Altern Med Rev. 2005 Sep;10(3):172-92.
Celiac disease develops from an autoimmune response to specific dietary grains that contain gluten. Diagnosis can be made based on the classical presentation of diarrhea, fatty stools, and abdominal bloating and cramping, as well as the presence of specific serum antibodies. In addition, gluten ingestion has increasingly been found to be associated with other conditions not usually correlated with gluten intolerance. The subsequent diversity of the clinical presentation in these cases can complicate decision-making and delay treatment initiation in conditions such as ataxia, headaches, arthritis, neuropathy, type 1 diabetes mellitus, and others. This review explores the etiology and pathology of celiac disease, presents support for the relationship between gluten and other diseases, and provides effective screening and treatment protocols. [Abstract/Link to Full Text]

Czap A
Color blind, or just plain blind?
Altern Med Rev. 2005 Jun;10(2):81-2. [Abstract/Link to Full Text]

L-arginine. Monograph.
Altern Med Rev. 2005 Jun;10(2):139-48. [Abstract/Link to Full Text]

Recent Articles in Journal of Physiology and Pharmacology

Konturek SJ, Konturek PC, Brzozowska I, Pawlik M, Sliwowski Z, Cze?nikiewicz-Guzik M, Kwiecie? S, Brzozowski T, Bubenik GA, Pawlik WW
Localization and biological activities of melatonin in intact and diseased gastrointestinal tract (GIT).
J Physiol Pharmacol. 2007 Sep;58(3):381-405.
Melatonin (MT), an indole formed enzymatically from L-trytophan (Trp), was first discovered in the bovine pineal gland in 1958 by Lerner et al. Melatonin is the most versatile and ubiquitous hormonal molecule produced not only in the pineal gland but also in various other tissues of invertebrates and vertebrates, particularly in the gastrointestinal tract (GIT). This review focuses on the localization, production, metabolism and the functions of MT in GIT and the duodenal unit (liver, biliary routes and pancreas), where multi-step biosynthetic pathways of this indole, similar to those in pinealocytes, have been identified. These biosynthetic steps of MT, including two major rate limiting enzymes; arylalkylamine-N-acetyltransferase (AA-NAT) and hydroxyindole-O-methyltransferase (HIOMT), transforming L-tryptophan (Trp), originally identified in pinealocytes, have been also detected in entero-endocrine (EE) cells of GIT, where this indole appears to act in endocrine, paracrine and/or luminal pathway directly or through G-protein coupled MT receptors. Studies of the distribution of MT in GIT mucosa showed that this indole is generated in GIT in much larger amounts than it is produced in the pineal gland. Melatonin acts in GIT, partly locally in paracrine fashion and is partly released into portal circulation, to be taken up by the liver. It is then metabolized and excreted with the bile to small bowel and finally returns to liver through entero-hepatic circulation. The production of MT by the pineal gland shows circadian rhythm with high night-time surge, especially at younger age, followed by the fall during the day-light time. As a highly lipophylic substance, MT reaches all body cells within minutes, thus, serving as a convenient circadian timing signal. Following pinealectomy, the light/dark cycle of plasma MT levels disappears, while its day-time blood concentration is maintained mainly due to its release from the GIT. According to our experience, after oral application of Trp, the plasma MT increases in dose-dependent manner both in intact and pinealectomized animals and humans, indicating that GIT but not the pineal gland is a source of this indole. In GIT MT exhibits a wide spectrum of activities such as circadian entrainment, antioxidant and free radicals scavenging activity, Melatonin (MT), an indole formed enzymatically from L-trytophan (Trp), was first discovered in the bovine pineal gland in 1958 by Lerner et al. Melatonin is the most versatile and ubiquitous hormonal molecule produced not only in the pineal gland but also in various other tissues of invertebrates and vertebrates, particularly in the gastrointestinal tract (GIT). This review focuses on the localization, production, metabolism and the functions of MT in GIT and the duodenal unit (liver, biliary routes and pancreas), where multi-step biosynthetic pathways of this indole, similar to those in pinealocytes, have been identified. These biosynthetic steps of MT, including two major rate limiting enzymes; arylalkylamine-N-acetyltransferase (AA-NAT) and hydroxyindole-O-methyltransferase (HIOMT), transforming L-tryptophan (Trp), originally identified in pinealocytes, have been also detected in entero-endocrine (EE) cells of GIT, where this indole appears to act in endocrine, paracrine and/or luminal pathway directly or through G-protein coupled MT receptors. Studies of the distribution of MT in GIT mucosa showed that this indole is generated in GIT in much larger amounts than it is produced in the pineal gland. Melatonin acts in GIT, partly locally in paracrine fashion and is partly released into portal circulation, to be taken up by the liver. It is then metabolized and excreted with the bile to small bowel and finally returns to liver through entero-hepatic circulation. The production of MT by the pineal gland shows circadian rhythm with high night-time surge, especially at younger age, followed by the fall during the day-light time. As a highly lipophylic substance, MT reaches all body cells within minutes, thus, serving as a convenient circadian timing signal. Following pinealectomy, the light/dark cycle of plasma MT levels disappears, while its day-time blood concentration is maintained mainly due to its release from the GIT. According to our experience, after oral application of Trp, the plasma MT increases in dose-dependent manner both in intact and pinealectomized animals and humans, indicating that GIT but not the pineal gland is a source of this indole. In GIT MT exhibits a wide spectrum of activities such as circadian entrainment, antioxidant and free radicals scavenging activity, cytoprotective, anti-inflammatory and healing efficacy of various GIT lesions such as esophagitis, gastritis, peptic ulcer, pancreatitis and colitis. This review concentrates on the generation and pathophysiological implication of MT in GIT and related organs. [Abstract/Link to Full Text]

Konturek SJ, Zayachkivska O, Havryluk XO, Brzozowski T, Sliwowski Z, Pawlik M, Konturek PC, Cze?nikiewicz-Guzik M, Gzhegotsky MR, Pawlik WW
Protective influence of melatonin against acute esophageal lesions involves prostaglandins, nitric oxide and sensory nerves.
J Physiol Pharmacol. 2007 Jun;58(2):361-77.
Melatonin (MT) is known to protect gastrointestinal mucosa against various types of injury but its effects on esophageal damage have not been studied. We examined the effects of MT on acute esophageal injury and the mechanism involved in the action of this indole. Acute esophageal lesions were induced by perfusion with acid-pepsin solution using tube inserted through the oral cavity into the mid of esophagus of anaesthetized rats with or without inhibition of prostaglandin (PG) generation by indomethacin (5 mg/kg/day), nitric oxide (NO) formation by N(G)-nitro-L-arginine (L-NNA, 20 mg/kg/day) or sensory nerves deactivation by capsaicin (125 mg/kg, sc). The esophageal injury was assessed by macroscopic score and histologic activity index. The esophageal mucosal blood flow (EBF) was determinated by H(2)-gas clearance method. The plasma TNF-alpha and nitrate/nitrite (NOx) levels and mucosal PGE(2) contents were assessed by immunoassays. Esophageal acid-pepsin perfusion induced noticeable esophageal mucosal injury as compared to perfusion with vehicle saline. The pretreatment with MT prevented significantly esophageal injury, raised EBF and mucosal content of PGE(2), while decreasing the levels of TNF-alpha. Inhibition of COX/PG and NOS/NO systems by indomethacin and L-NNA, respectively, or inactivation of sensory nerves by capsaicin, that manifested in further increase of esophageal injury, reduced the levels of EBF, markedly raised the levels TNF-alpha and reduced mucosal PGE(2), but the pretreatment with MT prevented significantly esophageal injury, improved EBF and raised mucosal PGE(2) contents. These studies suggest that MT can be considered as a novel esophagoprotector, acting, at least in part, through the COX/PG and NOS/NO systems and activation of sensory nerves. [Abstract/Link to Full Text]

Bojanowska E, Nowak A
Interactions between leptin and exendin-4, a glucagon-like peptide-1 agonist, in the regulation of food intake in the rat.
J Physiol Pharmacol. 2007 Jun;58(2):349-60.
Leptin interplays with other peptides to control feeding behaviour in humans and animals. Using exendin-4, an agonist of glucagon-like peptide-1, we investigated whether leptin modifies its effect on food intake in the rat. In the first series, exendin-4 alone (0.1, 2 or 10 microg per rat), leptin alone (0.1, 2, 10 or 100 microg per rat) or exendin-4 and leptin together (0.1 + 0.1, 2 + 2, 10 + 10, or 2 + 100 microg per rat, respectively) were injected once intraperitoneally. In the second series animals were injected either with exendin-4 (2 microg) alone, leptin (10 microg) alone, or leptin (10 microg) + exendin-4 (2 microg) daily for 5 subsequent days. At the lowest dose used, leptin and exendin-4 injected once together, but not separately, reduced significantly a 24-hour food intake. When used in higher doses, however, leptin did not change the exendin-4-dependent suppressory effect on food consumption. No significant differences in food intake were seen between rats treated repeatedly with exendin-4 only and animals injected with both drugs. Hence, leptin and exendin-4 may act additively to inhibit appetite when present in low concentrations while, at high leptin doses, this effect is abolished. The lack of synergistic effects of exendin-4 and high leptin concentrations on food intake may explain, at least in part, mechanisms responsible for leptin resistance in subjects with hyperleptinaemia. [Abstract/Link to Full Text]

Bugajski AJ, Zurowski D, Thor P, Gadek-Michalska A
Effect of subdiaphragmatic vagotomy and cholinergic agents in the hypothalamic-pituitary-adrenal axis activity.
J Physiol Pharmacol. 2007 Jun;58(2):335-47.
In the present study, we examined whether the vagus nerve is involved in mediating the stimulation of hypothalamic-pituitary-adrenal (HPA) axis by cholinergic muscarinic and nicotinic agonists, carbachol and nicotine. The site of HPA axis muscarinic stimulation was determined using peripheral (i.p.) and intracerebroventricular (i.c.v.) administration of carbachol, atropine sulphate (AtrS) and atropine hydrobromide (AtrBr). The i.p. carbachol-(0.5 mg/kg)-induced corticosterone response was significantly reduced by i.p. pretreatment with AtrBr (0.1 mg/kg), but was not diminished by i.c.v. AtrS (0.1 mug). The increase in corticosterone secretion induced by i.c.v. carbachol (2 microg) was totally abolished by i.c.v. pretreatment with AtrS (0.1 microg) but was not altered by i.p. AtrBr. Subdiaphragmatic vagotomy performed 2 weeks earlier substantially decreased the i.p. carbachol (0.2 mg/kg)-induced ACTH response and markedly augmented ACTH and corticosterone response to a higher dose of carbachol (0.5 mg/kg) in comparison with the responses in sham operated rats. Vagotomy abolished the stimulatory effect of i.p. nicotine in a low dose (1 mg/kg) on ACTH and corticosterone secretion; the ACTH response to higher dose (2.5 mg/kg) was considerably reduced, while corticosterone response remained unaffected. These results suggest that carbachol given i.c.v. evokes considerable corticosterone response by stimulation of central cholinergic muscarinic receptors. A major part of the i.p. carbachol-induced corticosterone secretion results from peripheral cholinergic muscarinic receptor stimulation. Subdiaphragmatic vagotomy moderately intensified the carbachol-induced ACTH and corticosterone secretion. Vagotomy significantly reduced the nicotine-induced ACTH secretion, possibly by the involvement of vagal afferents. The nicotine-induced corticosterone secretion is not exclusively regulated by circulating ACTH but by various intra-adrenal regulatory components. [Abstract/Link to Full Text]

Karczewska J, Martyniec L, Dzierzko G, Stepi?ski J, Angielski S
The relationship between constitutive ATP release and its extracellular metabolism in isolated rat kidney glomeruli.
J Physiol Pharmacol. 2007 Jun;58(2):321-33.
ATP and adenosine are important extracellular regulators of glomerular functions. In this study, ATP release from glomeruli suspension and its extracellular metabolism were investigated. Basal extraglomerular ATP concentration (1nM) increased several fold during inhibition of ecto-ATPase activity, reflecting the basal ATP release rate. Mechanical perturbation increased the amounts of ATP released from glomeruli. ATP added to glomeruli was almost completely degraded within 20 minutes. In that time, AMP was the main product of extracellular ATP metabolism. Significant accumulation of AMP was observed after 5 min (194 +/-16 microM) and 20 min (271 +/-11 microM), whereas at the same time concentration of adenosine was only 10 muM. A competitive inhibitor of ecto-5-nucleotidase alpha-beta-methylene-ADP (AOPCP), decreased extraglomerular ATP and adenosine concentration by 80% and 50%, respectively. Similarly, AMP (100 microM) also markedly reduced extraglomerular ATP accumulation, whereas IMP, its deamination product, was not effective. P1, P5-diadenosine pentaphosphate (Ap5A) - an inhibitor of ecto-adenylate kinase prevented significantly the disappearance of ATP from extraglomerular media caused by AMP. These findings demonstrate that the decrease in extracellular ATP concentration observed after addition of AOPCP or AMP is caused by the presence of ecto-adenylate kinase activity in the glomeruli. The enzyme catalyses reversible reaction 2ADP<->ATP+AMP, and a rise in the AMP concentration can lead to fall in ATP level. The present study provides evidence the extraglomerular accumulation of ATP reflects both release of ATP from glomeruli cells and its metabolism by ecto-enzymes. Our data suggest that AMP, produced from ATP in the Bowman's capsular space, might plays a dual role as a substrate for ecto-adenylate kinase and ecto-nucleotidase reactions being responsible for the regulation of intracapsular ATP and adenosine concentration. We conclude that AMP degrading and converting ecto-enzymes effectively determine the balance between ATP and adenosine concentration and thus the activation of P2 and/or adenosine receptors. [Abstract/Link to Full Text]

Warzecha Z, Dembi?ski A, Ceranowicz P, Dembi?ski M, Cieszkowski J, Ku?nierz-Cabala B, Naskalski JW, Jaworek J, Konturek SJ, Pawlik WW, Tomaszewska R
Influence of ischemic preconditioning on blood coagulation, fibrinolytic activity and pancreatic repair in the course of caerulein-induced acute pancreatitis in rats.
J Physiol Pharmacol. 2007 Jun;58(2):303-19.
Previous studies have shown that ischemic preconditioning protects several organs, including the pancreas, from ischemia/reperfusion-induced injury. The aim of the investigation was to determine whether ischemic preconditioning affects the course edematous pancreatitis. METHODS: In rats, ischemic preconditioning was performed by short-term clamping the celiac artery. Acute pancreatitis was induced by caerulein. The severity of acute pancreatitis was evaluated between the first and tenth day of inflammation. RESULTS: Ischemic preconditioning applied alone caused a mild pancreatic damage. Combination of ischemic preconditioning with caerulein attenuated the severity of pancreatitis in histological examination and reduced the pancreatitis-evoked increase in plasma lipase and pro-inflammatory interleukin-1beta. This effect was associated with an increase in plasma level of anti-inflammatory interleukin-10 and partial reversion of the pancreatitis-evoked drop in pancreatic DNA synthesis and pancreatic blood flow. In secretory studies, ischemic preconditioning in combination with induction of acute pancreatitis attenuated the pancreatitis-evoked decrease in secretory reactivity of isolated pancreatic acini to stimulation by caerulein. In the initial period of acute pancreatitis, ischemic preconditioning alone and in combination with caerulein-induced acute pancreatitis prolonged the activated partial thromboplastin time (APTT), increased plasma level of D-dimer and shortened the euglobulin clot lysis time. The protective effect of ischemic preconditioning was observed during entire time of experiment and led to acceleration of pancreatic regeneration. CONCLUSIONS: Ischemic preconditioning reduces the severity of caerulein-induced pancreatitis and accelerates pancreatic repair; and this effect is related to the activation of fibrinolysis and reduction of inflammatory process. [Abstract/Link to Full Text]

Jaworek J, Nawrot-Porabka K, Leja-Szpak A, Szklarczyk J, Macko M, Bonior J, Stachura J, Konturek SJ, Pawlik WW
Exposition of newborn rats to bacterial endotoxin impairs pancreatic enzyme secretion at adult age.
J Physiol Pharmacol. 2007 Jun;58(2):287-302.
Lipopolysaccharide (LPS, endotoxin) is the component of the cellular wall of Gram negative bacteria. Endotoxemia (sepsis) could produce multiorgan failure and in the early period of life LPS are responsible for the changes of metabolism and for the reduction of protein synthesis. The influence of neonatal endotoxemia on the pancreas at adults has not been investigated yet. The aim of this study was to assess the pancreatic exocrine function in the adult rats which have been subjected, in the neonatal period of life, to chronic LPS pretreatment. LPS from E. coli or S. typhi at doses of 5, 10 or 15 mg/kg-day was administered intraperitoneally (i.p.) to the suckling rats (30 g) during 5 consecutive days. Three months later these animals (300 g) were equipped with pancreato-biliary fistulae for the in vivo secretory study. Amylase release from isolated pancreatic acini obtained from these rats was also assessed. Pancreatic tissue samples were taken for histological assessment and for the determination of gene expression for CCK1 receptor by RT-PCR. Pancreatic amylase secretions stimulated by caerulein or by diversion of pancreatic-biliary juice to the exterior (DBPJ) was significantly, and dose-dependently reduced in the adult rats which have been subjected in infancy to chronic pretreatment with LPS from E. coli or S. typhi, as compared to the untreated control. In these animals basal secretion was unaffected. In the rats pretreated with LPS in the suckling period of life caerulein-induced amylase release from isolated pancreatic acini was significantly decreased, as compared to the untreated with LPS control. This was accompanied by dose-dependent reduction of mRNA signal for CCK1 receptor on pancreatic acini. Neonatal endotoxemia failed to affect significantly pancreatic morphology as well as plasma amylase level in the adult rats. We conclude that neonatal endotoxemia reduces gene expression for CCK1 receptor and could produce impairment of the exocrine pancreatic function at adult age. [Abstract/Link to Full Text]

Santiago-Mejia J, Ventura-Martinez R, Gomez C, Parra-Gamez L, Gonzales-Rios J, Rodriguez R
Polyviewed expression of the altered contractility of the guinea-pig ileum after ischemia in situ and superfusion in vitro.
J Physiol Pharmacol. 2007 Jun;58(2):275-85.
This study deals with the polyviewed expression of the altered contractility of the isolated ileum of the guinea-pig after ischemia/superfusion (I/S). Intestinal ischemia was produced by clamping the superior mesenteric artery for 40, 80 or 160 min. Ischemic and non-ischemic segments taken from the same guinea-pig were mounted for tension recording in organ baths and superfused (120 min) with an oxygenated Krebs-bicarbonate solution. Data were analyzed by means of the Polyview System software, which allows detecting simultaneously several events of one response. Histopathological changes in myenteric neurons were also examined. We found that ischemia in situ followed by superfusion in vitro (reoxygenation) severely reduces the spontaneous intestine contractile activity, and significantly decreases the maximal contractile response to ACh and to electrical field stimulation (EFS), the maximal rate of tension, and the sensitivity of the tissue to EFS. In addition, these ischemic intestines respond with a long-lasting contracture when electrical stimulation was started at supramaximal voltage. Functional alterations were time dependent. Neurons exhibited features of necrosis. These results provide clear evidence of detrimental effects of I/S on intestine contractile function. Digital analysis allows quantification of additional parameters important for evaluation of functional changes after I/S and of the degree of neuroprotection. [Abstract/Link to Full Text]

Zoladz JA, Szkutnik Z, Majerczak J, Duda K, Pedersen PK
Non-linear relationship between oxygen uptake and power output in the Astrand nomogram-old data revisited.
J Physiol Pharmacol. 2007 Jun;58(2):265-73.
For the last decade there have been considerable discussion concerning the linearity / non-linearity of the oxygen uptake (V(O2)) - power output (W) relationship with strong experimental evidence of non-linearity provided mainly by breath-by-breath measurements. In this study, we attempted to answer the question whether the V(O2) - W relationship in the Astrand nomogram, as presented in the Textbook of Work Physiology, P.-O. Astrand et al. (2003), page 281, based on the Douglas bag method, is indeed linear, as stated by the authors before, or if a change point in V(O2), described by Zoladz et al. (1998) Eur J Appl Physiol 78: 369-377, can possibly be detected in those data. The V(O2) - W data were taken from the Astrand nomogram referenced above and from the Table 9.5 on page 282 in the same reference and tested for the presence of the change point in V(O2), using our two-phase model (see the reference above). In the first phase, a linear V(O2) - W relationship was assumed, whereas in the second one (above the so-called change point) an additional increase in V(O2) above the values expected from the linear model was allowed. It was found that in the data taken from the Astrand nomogram (data for men), as well as in the data taken from the Table 9.5, statistically significant change points in V(O2) were present at the power output of 150 W. The documentation of the presence of a change point in the V(O2) - W relationship in the Astrand data provides further evidence for the existence of a non-linearity in the V(O2) - W relationship in incremental exercise tests of humans, also in V(O2) data based upon the Douglas bag method. [Abstract/Link to Full Text]

Nowakowska E, Kus K, Czubak A, Glowacka D, Matschay A
Some behavioural effects of carbamazepine - comparison with haloperidol.
J Physiol Pharmacol. 2007 Jun;58(2):253-64.
The experiments presented in this paper aimed to investigate the influence of atypical antiepileptic drug carbamazepine (CBZ, CAS 298-46-4) classified also as normothymic drug on spatial memory in Morris water maze test and anxiolytic effect in two-compartment exploratory test in rats. The study also investigated the probably occurring side effects (measuring cataleptic activity and motor coordination) following single and chronic administration of CBZ compared to haloperidol (HAL, CAS 52-86-8), a conventional antipsychotic. All the tests were carried out on male Wistar rats. CBZ 30 mg/kg was administered orally 60 min before the tests and HAL 0.15 mg/kg was administered orally 60 min before the tests. In the Morris test memory improvement only after chronic administration of CBZ on the 7 and 14 day of treatment was observed, whereas after 14 days of HAL treatment spatial memory impairment was noted. In the two-compartment exploratory test 30 mg/kg of CBZ had an anxiolytic effect after 7 and 14 days of treatment, whereas HAL did not show anxiolytic effect after single and chronic treatment. CBZ did not induce catalepsy after single as well as chronic administration. HAL evoked a strong cataleptic effect both after acute and chronic treatment. CBZ had no impact on motor coordination in the chimney test and HAL disturbed motor coordination in rats after single as well as chronic administration. CBZ may be an useful normothymic drug using in bipolar affective disorder treatment with co-occurred anxiety and cognitive deficits. The lack of significant side effects of CBZ may be an alternative way of treatment in comparison with older drugs, such as lithium carbonate. [Abstract/Link to Full Text]

Wasik A, Roma?ska I, Antkiewicz-Michaluk L
The effect of an endogenous compound 1-methyl-1,2,3,4,-tetrahydroisoquinoline on morphine-induced analgesia, dependence and neurochemical changes in dopamine metabolism in rat brain structures.
J Physiol Pharmacol. 2007 Jun;58(2):235-52.
1,2,3,4-Tetrahydroisoquinolines, among them the most interesting neuroprotective substance, an inhibitor of MAO, 1-methyl-1,2,3,4-tetrahydroisoquinoline (1MeTIQ), are endogenous compounds present in the central nervous system of mammals and humans. In this study, we investigated the effect of 1MeTIQ on morphine-induced analgesia, tolerance and abstinence syndrome as well as its effect on morphine-induced changes in dopamine metabolism in rat brain structures (nucleus accumbens, striatum, substantia nigra) using HPLC methodology. The experiments were carried out on male Wistar rats. Morphine analgesia was measured in the "hot-plate" test. To induce tolerance, morphine was given chronically (20 mg/kg i.p.) alone or following 1MeTIQ (50 mg/kg i.p.) injection. The development of dependence was assessed in the naloxone (2 mg/kg i.p.) precipitation test, after 10 days of morphine administration. The behavioral studies have shown that an endogenous compound, 1MeTIQ produced strong potentiation of morphine analgesia, prevented the development of morphine tolerance and inhibited expression of morphine abstinence syndrome in morphine-dependent rats. In neurochemical studies, we have demonstrated that 1MeTIQ antagonized morphine-induced changes in dopamine metabolism observed in rat brain structures. The main finding of this study was demonstration for the first time of an anti-abuse effect of an endogenous compound, 1MeTIQ, and its efficiency in counteracting morphine-induced addiction in the way useful from clinical point of view. The obtained results suggested a possibility of clinical application of 1MeTIQ in morphine addiction. [Abstract/Link to Full Text]

Rogóz Z, Skuza G, Legutko B
Repeated co-treatment with imipramine and amantadine induces hippocampal brain-derived neurotrophic factor gene expression in rats.
J Physiol Pharmacol. 2007 Jun;58(2):219-34.
The problem of drug-resistant depression indicates a strong need for alternative antidepressant therapies. In our earlier papers we described synergistic, antidepressant-like effects of a combination of imipramine (IMI) and amantadine (AMA) in the forced swimming test in rats, an animal model of depression. Moreover, preliminary clinical data showed that the above-mentioned combination had beneficial effects in treatment-resistant patients. In addition, a number of studies predicted a role of the brain-derived neurotrophic factor (BDNF) in the mechanism of action of antidepressant drugs (ADs). Since the most potent effect of ADs on BDNF gene expression was found after prolonged treatment, in the present study we investigated the influence of repeated treatment with IMI (5 or 10 mg/kg) and AMA (10 mg/kg), given separately or jointly (twice daily for 14 day), on mRNA level (the Northern blot) in the hippocampus and cerebral cortex. The experiment was carried out on male Wistar rats. The tissue for biochemical assays was dissected 24 h after the last dose of IMI and AMA. We also studied the effect of repeated treatment with IMI and AMA on the action of 5-HT(1A)- and 5-HT(2A) receptor agonists (8-OH-DPAT and (+/-)DOI, respectively) in behavioral tests. The obtained results showed that in the hippocampus IMI (10 mg/kg), and in the cerebral cortex IMI (5 and 10 mg/kg) and AMA (10 mg/kg) significantly elevated BDNF mRNA level. Joint administration of IMI (5 or 10 mg/kg) and AMA (10 mg/kg) induced a more potent increase BDNF gene expression in the hippocampus (but not in cerebral cortex) and either inhibited the behavioral syndrome induced by (+/-)DOI or did not change the action of 8-OH-DPAT (compared to treatment with either drug alone). The obtained results suggest that the enhancement of BDNF gene expression may be essential for the therapeutic effect of co-administration of IMI and AMA to drug-resistant depressed patients, and that among other mechanisms, 5-HT(2A) receptors possibly play some role in this effect. [Abstract/Link to Full Text]

Pierzchalska M, Soja J, Wo? M, Szabó Z, Nizankowska-Mogielnicka E, Sanak M, Szczeklik A
Deficiency of cyclooxygenases transcripts in cultured primary bronchial epithelial cells of aspirin-sensitive asthmatics.
J Physiol Pharmacol. 2007 Jun;58(2):207-18.
BACKGROUND: Airway function is actively regulated by epithelium through generating PGE(2), the production of which depends on cyclooxygeneses (COX-1 and COX-2). Analysis of bronchial biopsies and bronchial epithelial cells in culture conducted so far gave conflicting results of expression pattern of these enzymes in healthy subjects and asthmatics patients, with and without aspirin hypersensitivity. OBJECTIVE: Our aim was to investigate the expression of COX-1 and COX-2 mRNA in primary human bronchial epithelial cells (HBEC) isolated from asthmatics and non-asthmatics. METHODS: We isolated HBEC from bronchial brushing preparations taken during bronchoscopy of 10 non-asthmatics (NA), 8 aspirin-tolerant asthmatics (ATA) and 9 aspirin-intolerant asthmatics (AIA). HBEC were cultured in serum free medium until 80% confluent. Total cellular RNA was isolated and reversed transcribed using oligo(dT)(15) primers. Real time PCR was performed with primers to COX-1, COX-2, GAPDH and beta-actin in the presence of SYBR green dye. The cycle threshold (C(T)) for COX-1 or COX-2 was normalized using beta-actin and GAPDH as the internal standards. RESULTS: Not only COX-1 but also COX-2 mRNA were expressed by HBEC without any proinflammatory stimulation. We detected the smallest amount of COX-1 mRNA in the AIA group. The same trend was observed for COX-2 mRNA, though it didn't reach the statistical significance. We also analysed the relationship between DeltaC(TCOX-1) to DeltaC(TCOX-2) by calculating the difference DeltaDeltaC(TCOX-1-COX-2). This analysis revealed that AIA group can be characterized by relatively smallest COX-1 mRNA expression in comparison to COX-2. There is a strong positive correlation between C(TCOX1) and C(TCOX2) in NA group (r=0.85; p< 0.001). In both groups of asthmatics this correlation is absent (ATA - r=0.5, p>0.1; AIA - r=0.43, p>0.1). CONCLUSIONS: Cyclooxygeneases transcripts expression is altered in HBEC derived from the asthmatic patients, and this phenomenon is pronounced in case of aspirin hypersensitivity. [Abstract/Link to Full Text]

Jin L, Caldwell RB, Li-Masters T, Caldwell RW
Homocysteine induces endothelial dysfunction via inhibition of arginine transport.
J Physiol Pharmacol. 2007 Jun;58(2):191-206.
Hyperhomocysteinemia is an independent risk factor for cardiovascular diseases. High levels of plasma homocysteine (HCY) increase oxidative stress and reduce endothelial-dependent relaxation. We determined whether hyperhomocysteinemia-induced endothelial dysfunction is mediated through inhibition of cellular transport of L-arginine. In endothelial cells, HCY had a biphasic effect on arginine transport. HCY treatment for 6 hr increased L-arginine uptake by 34%; however, uptake was decreased by 25% after 24 h. HCY caused membrane hyperpolarization during both 6 and 24 h incubation periods, indicating that the negative charge facilitating arginine uptake was maintained. HCY significantly reduced expression of cellular arginine transporter protein (CAT-1) after 24 h treatment; whereas endothelial nitric oxide synthase (eNOS) protein levels and basal eNOS activity were not altered. Nevertheless, nitric oxide (NO) formation was significantly decreased. The antioxidant ascorbic acid prevented the effect of HCY on arginine transport. HCY induced formation of the peroxynitrite biomarker nitrotyrosine, which was blocked by supplemental L-arginine. HCY treatment of aortic rings caused decreased vasorelaxation to acetylcholine, which was prevented by supplemental arginine. In conclusion, HCY decreased NO formation and induced endothelial dysfunction without altering protein level or basal activity of eNOS, but through decreases in function and protein expression of the CAT-1 transporter. Reduced arginine supply may lead to eNOS uncoupling and generation of superoxide, contributing to HCY-induced oxidative stress. [Abstract/Link to Full Text]

Misiolek M, Marek B, Namyslowski G, Scierski W, Zwirska-Korczala K, Kazmierczak-Zagorska Z, Kajdaniuk D, Misiolek H
Sleep apnea syndrome and snoring in patients with hypothyroidism with relation to overweight.
J Physiol Pharmacol. 2007 Mar;58 Suppl 177-85.
The relation between snoring and obstructive sleep apnea as well as hypothyroidism is the object of interest of many authors. The respiratory disturbances during sleep are often observed in patients suffering from hypothyroidism. The relation of snoring to overweight in those patients has not been taken into account. The aim of the study was to evaluate the relations between hypothyroidism and quantitative and qualitative respiratory disturbances during sleep. Additional aim was to establish the relations of sleep apnea syndrome, snoring, hypothyroidism and overweight. The subjects included 15 patients (11 females and 4 males) aged from 28 to 73 (mean 50.3) suffering from hypothyroidism. All of them underwent thyroid testing before and after the hormonal treatment. TSH and fT4 concentrations were determined. At the same time the sleep assessment (PolyMESAM) was performed twice. Data were obtained from sleep studies and questionnaires (Epworth sleepiness scale). After the thyroid hormones stabilization significant decrease of snoring severity was observed. On the contrary, the respiratory disturbance index (RDI), desaturation index (DI), the lowest saturation (LSAT) did not change significantly, however, the Epworth scale score showed significant improvement. The correlations showed the strong relation between loud snoring and TSH (r=0.73, p<0.01) and fT4 (r=-0.66, p<0.003) concentrations before the treatment. The analysis showed no correlation between body mass (BMI) and snoring. The hormonal stabilization in patients suffering from hypothyroidism causes improvement in snoring severity. Based on our investigation the relationship between hypothyroidism and severity of snoring and excessive daytime somnolence was confirmed. It indicates a possible connection between hypothyroidism and upper airway resistance syndrome. [Abstract/Link to Full Text]

Harsch IA, Bergmann T, Koebnick C, Wiedmann R, Ruderich F, Hahn EG, Konturek PC
Adiponectin, resistin and subclinical inflammation--the metabolic burden in Launois Bensaude Syndrome, a rare form of obesity.
J Physiol Pharmacol. 2007 Mar;58 Suppl 165-76.
The aim of the study was to investigate, whether the degree of metabolic risk factors for atherosclerotic complications in a very rare kind of obesity, the Multiple Symmetrical Lipomatosis, also known as the Launois-Bensaude Syndrome (LBS), are comparable or different from "simple" truncal obesity. 10 patients with LBS (Body mass index 34.4 +/- 1.8 kg/m(2), age: 62 +/- 3 yrs) were compared with 19 BMI - matched patients with "simple" truncal obesity and obstructive sleep apnoea syndrome (OSAS) and 20 BMI- matched patients with "simple" truncal obesity without OSAS. Markers of subclinical inflammation and thrombocyte activation (sCD62p = soluble p-selectin, highly sensitive C-Reactive protein = CRP, Interleukin-6 = IL-6, ICAM-1 = Intracellular Adhesion Molecule-1, Vascular Cell Adhesion Molecule = VCAM -1, leptin), as well as adiponectin and resistin were studied. The prevalence of atherogenic risk factors as hypertension (80%), type 2 diabetes (30%), OSAS (50%), smoking (30%) and alcohol abuse (80%) was high in the (obese) LBS group. The markers of subclinical inflammation and thrombocyte activation showed an indifferent picture with lower levels of circulating IL-6 and sCD62p, comparable CRP and higher ICAM-1 and VCAM-1 than in controls. Leptin and adiponectin were higher than in controls. However, the accumulation of "classic" cardiovascular risk factors in the LBS group was well reflected by the presence of symptomatic cardiovascular disease in 3 of the 10 LBS patients, putting LBS patients - if obese - at an atherosclerotic risk at least comparable to obese persons. [Abstract/Link to Full Text]

Zwirska-Korczala K, Adamczyk-Sowa M, Sowa P, Pilc K, Suchanek R, Pierzchala K, Namyslowski G, Misiolek M, Sodowski K, Kato I, Kuwahara A, Zabielski R
Role of leptin, ghrelin, angiotensin II and orexins in 3T3 L1 preadipocyte cells proliferation and oxidative metabolism.
J Physiol Pharmacol. 2007 Mar;58 Suppl 153-64.
There is now growing evidence that the reactive oxygen species have an influence on proliferation and antioxidative status of various cell types. The aim of the study was to investigate the effects of different concentrations of leptin, ghrelin, angiotensin II and orexins on proliferation, culture medium malondialdehyde (MDA) levels and antioxidative enzymes activities: superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and catalase (CAT) in 3T3 L1 preadipocytes cell culture. Cell proliferation was measured using [(3)H]tymidine incorporation. In 3T3-L1 cells leptin caused a significant reduction in proliferation (by 36%) compared to control. Ghrelin increased preadipocyte proliferation, and the effect was stronger in higher dose (by 39%), while proproliferatory effect of angiotensin II was stronger in lower doses (by 47%). All used doses of orexin A significantly increased 3T3 L1 cell proliferation (from 21% to 160%), while orexin B caused a marked reduction (from 35% to 70%) of this proliferation. The effects of both orexins were dose-dependent. Leptin and ghrelin increased activity of SOD, CAT, GSH-Px and decreased level of MDA. Angiotensin II treatment stimulated only SOD and CAT activities. Influence of orexins was different on various enzymes. Orexin A increased MDA levels, while orexin B caused a marked decrease in MDA levels. Our results strongly suggest the effects of appetite affecting hormones such as leptin and ghrelin on proliferation and antioxidative enzyme activities of preadipocyte cell lines. Orexin A was found to be the most efficient proliferative-signalling hormone, while orexin B revealed the most significant inhibitory effect on preadipocytes proliferation. [Abstract/Link to Full Text]

Sodowski K, Zwirska-Korczala K, Kuka D, Kukla M, Budziszewska P, Czuba B, W?och A, Cnota W, Biela?ski W, Brzozowski T, Rehfeld JF, Zdun R, Konturek PC
Basal and postprandial gut peptides affecting food intake in lean and obese pregnant women.
J Physiol Pharmacol. 2007 Mar;58 Suppl 137-52.
Maternal obesity has been reported as a risk factor for various maternal and fetal complications. The aim of the present study was to examine the patterns of basal and postprandial plasma concentrations of certain gut hormones affecting food intake such as acylated ghrelin, peptide YY(3-36) (PYY(3-36)), cholecystokinin (CCK), insulin and glucose in pregnant women with varying body mass gain during physiological pregnancy. The study included 34 women with singleton pregnancies in the 2(nd) trimester of gestation. The examined pregnant women were divided into 4 groups; I. control pregnancy (CP) with weight gain below 0.5 kg/week; II. overweight low weight gain <1 kg/week (OLWG), III. overweight high weight gain >1 kg/week (OHWG); morbidly obese pregnant with weight gain >1.5 kg/week (MOP). The basal acylated-ghrelin levels in MOP subjects were significantly higher than those in CP and no usual suppression of acylated ghrelin after the meal observed in CP as well as in OLWG and OHWG was found in MOP women. Basal PYY(3-36) plasma levels were similar in CP, OLWG and OHWG but in MOP was significantly reduced and no significant increase in hormone level, typically observed in CP, was detected after a meal in overweight or obese women studied. The fasting CCK and C-reactive protein (CRP) levels in MOP subjects were significantly higher than those in CP and other overweight women. In conclusion, we found that pregnant women with overweight and obesity exhibit significant changes in fasting and postprandial gut hormones affecting food intake such as acylated ghrelin, PYY(3-36) and CCK as well as in CRP and these changes might contribute, at least in part, the development of obesity in pregnancy. [Abstract/Link to Full Text]

Zwirska-Korczala K, Konturek SJ, Sodowski M, Wylezol M, Kuka D, Sowa P, Adamczyk-Sowa M, Kukla M, Berdowska A, Rehfeld JF, Bielanski W, Brzozowski T
Basal and postprandial plasma levels of PPY, ghrelin, cholecystokinin, gastrin and insulin in women with moderate and morbid obesity and metabolic syndrome.
J Physiol Pharmacol. 2007 Mar;58 Suppl 113-35.
Metabolic syndrome (MS), defined as central obesity, hyperinsulinemia, insulin resistance, hypertension, dyslipidemia and glucose intolerance, has been associated with inflammatory biomarkers and cardiovascular diseases. This study was carried out on three groups of women; lean controls, moderately obese with MS (OB-MS) and morbidly obese with MS (MOB-MS). The main objectives were: 1. to analyze the plasma levels of total and acylated ghrelin, peptide YY(3-36) (PYY(3-36)), cholecystokinin (CCK), gastrin and insulin levels under basal conditions and in response to a standard mixed meal, and 2. to elucidate the relationship between the plasma levels of these gut peptides and metabolic syndrome parameters. Plasma levels of the gut hormones were measured by radioimmunoassays at time 0 just before the meal and at 30, 60 and 120 min after a meal ingestion. Traditional lipid profile and high-sensitivity C reactive protein (hs-CRP), the strongest biomarker of inflammation were also determined in OB-MS and MOB-MS. When compared to OB-MS, MOB-MS exhibited much higher anthropometric parameters such as waist circumference, higher fat mass and higher plasma levels of low density lipoprotein-cholesterol (LDL-C) and hs-CRP. Both these obese groups revealed significantly higher values of body mass index (BMI), fat mass, total cholesterol (TC), LDL-C, fasting glucose, fasting insulin, insulin resistance (IR) calculated from homeostatic model assessment (HOMA) and hs-CRP compared to the values recorded in lean subjects. Fasting PYY(3-36) level was lower, while fasting acylated ghrelin was higher in MOB-MS than in OB-MS. Plasma total and acylated ghrelin levels were significantly lower in OB-MS compared to lean women. In MOB-MS women the fasting PYY(3-36) levels were lower compared to lean controls and OB-MS, whilst postprandially in both OB-MS and MOB-MS, it was much lower than in lean women. The fasting plasma levels of total and acylated ghrelin and their postprandial decrease were significantly smaller in both obese groups compared to lean subjects. Plasma hs-CRP levels correlated positively with BMI, waist circumference, fat mass, fasting glucose, HOMA IR and fasting active ghrelin, whilst it negatively correlated with plasma fasting and total ghrelin. Moreover, plasma fasting acylated ghrelin correlated positively with fat mass. Fasting total ghrelin correlated positively with BMI, HDL-C and negatively with HOMA IR. We conclude that MS features of obesity are closely related to fasting and postprandial alterations of concentrations of PYY(3-36), CCK and ghrelin, suggesting that determination of gut hormones controlling food intake might be considered as a valuable tool to assess the progression of MS to comorbidities of obesity. [Abstract/Link to Full Text]

Bugajski AJ, Gil K, Ziomber A, Zurowski D, Zaraska W, Thor PJ
Effect of long-term vagal stimulation on food intake and body weight during diet induced obesity in rats.
J Physiol Pharmacol. 2007 Mar;58 Suppl 15-12.
Regulation of food intake and body weight is accomplished by several mechanisms. CNS receives information from periphery and modifies food intake mainly by vagal nerves that provide the major neuroanatomical link between gastrointestinal sites stimulated during food intake and CNS sites that control feeding behavior and metabolism. Gastric mechanoreceptors and jejunal chemoreceptors activated by food or vagal nerve stimulation (VNS), which mimic the physiological input, suppress feeding within short-term regulation. Our research was aimed on determination the role of electrical VNS in long-term control of food intake and body weight in diet induced obesity fed rats. Food intake, body weight and epididymal fat pad were assessed in male Wistar rats divided into three groups (controls vs. VNS). Rats were implanted with microchip and kept during the whole study (100 days) on diet induced obesity. Vagal nerve was stimulated by electrical rectangular pulses duration 10 ms, amplitude 200 mV, frequency 0.05 Hz generated by microchip. In control group surgery produced no significant changes in meal size and body weight gain as compared to intact group. In contrast, significantly decreased epididymal fat pad weight, decreased meal size with effect on decreased weight gain was observed in VNS rats. Data support theory that VNS can increase vagal afferent signal conduct to CNS and mimics the satiety signals leading to reduce food intake and body weight gain. [Abstract/Link to Full Text]

Pawlikowski M, Lawnicka H, Pisarek H, Kunert-Radek J, Radek M, Culler MD
Effects of somatostatin-14 and the receptor-specific somatostatin analogs on chromogranin A and alpha-subunit (alpha-SU) release from "clinically nonfunctioning" pituitary adenoma cells incubated in vitro.
J Physiol Pharmacol. 2007 Mar;58(1):179-88.
The aim of the study was to examine the effect of somatostatin (SST) and its analogs on the release of chromogranin A (CgA) and alpha-subunit (alpha-SU) from clinically non-functioning pituitary adenomas incubated in vitro. Seven pituitary macroadenomas surgically removed were investigated. All of the tumors were diagnosed before surgery as non-functioning, but they expressed either gonadotropins or their subunits as detected by immunohistochemistry. Two tumors additionally expressed prolactin and growth hormone. All adenomas also expressed chromogranin A (CgA) and at least 3 of 5 subtypes of somatostatin receptors. The cells isolated from the examined tumors were exposed in vitro to either native SST-14 or the following receptor-specific SST analogs: BIM-23926 (agonist of sst1 receptor), BIM-23120 (agonist of sst2 receptor), BIM-23206 (agonist of sst5 receptor) and BIM23A387 (somatostatin/dopamine chimera). The concentration of CgA was measured by means of ELISA method and of alpha-SU was measured by an immunoradiometric method. It was found that the exposure on SST-14 resulted in the decrease of CgA and alpha-SU release from tumor cells in majority of samples, and the effect on CgA was positively correlated with the expression of sst3 and also with the sst2A/sst2B expressions ratio. The inhibitory effect of SST-14 on CgA and alpha-SU seems also to correlate negatively with the expression of sst2B. CgA inhibition also correlates positively with sst5 expression. Among the other compounds studied, only the sst2 agonist decreased the release in all the investigated samples. The remaining substances (agonists of sst1 and sst5 and SST/DA chimera) produced the divergent changes (increased or decreased release, depending on the sample). The data suggest that the inhibition of CgA (and possibly of alpha-SU) release by SST is mediated via subtypes sst2A, sst3 and sst5, whereas sst2B subtype may induce the opposite effect. [Abstract/Link to Full Text]

Mucha K, Foroncewicz B, Koziak K, Czarkowska-Paczek B, Paczek L
The effects of indomethacin on angiogenic factors mRNA expression in renal cortex of healthy rats.
J Physiol Pharmacol. 2007 Mar;58(1):165-78.
Indomethacin is a nonsteroidal anti-inflammatory drug used frequently to control chronic or temporary pain. In the kidney, indomethacin decreases medullary and cortical perfusion, resulting in hypoxia. Kidney hypoxia has many effects, including changes in gene expression, and is a strong stimulus for angiogenesis. Other angiogenic factors include vascular endothelial growth factor (VEGF), basic fibroblast growth factor (FGF-2), transforming growth factor beta 1 (TGFbeta1), and platelet-derived growth factor (PDGF). Our goal was to examine the influence of indomethacin on mRNA expression of these factors and their selected receptors in the renal cortex of healthy rats. Groups of 8 healthy, male, six-week-old Wistar rats received either indomethacin (5 mg/kg/day) or placebo orally for three months. RNA from renal cortex biopsies was analyzed by real-time polymerase chain reaction to quantify the mRNA levels of each cytokine. We observed significantly higher mRNA levels for VEGF (1.73-fold), FGF-2 (5.6-fold) and TGFbeta receptor III (2.93-fold), PDGF receptor alpha (2.93-fold) and receptor beta (2.91-fold) in rats receiving indomethacin compared to rats given placebo (p < 0.05). Amounts of mRNA for TGFbeta1, PDGF, FGF receptors 1 and 2 and TGFbeta receptor I did not differ between analysed groups. Our data indicates that indomethacin may regulate the expression of potent angiogenic factors VEGF and FGF-2. [Abstract/Link to Full Text]

Grzelec-Mojzesowicz M, Sadowski J
Renal tissue NO and intrarenal haemodynamics during experimental variations of NO content in anaesthetised rats.
J Physiol Pharmacol. 2007 Mar;58(1):149-63.
Direct renal nitric oxide (NO) measurements were infrequent and no simultaneous measurements of renal cortical and medullary NO and local perfusion. Large-surface NO electrodes were placed in renal cortex and medulla of anaesthetised rats; simultaneously, renal blood flow (RBF, index of cortical perfusion) and medullary laser-Doppler flux (MBF) were determined. NO synthase inhibitors: nonselective (L-NAME) or selective for neuronal NOS (nNOS) (S-methyl-thiocitrulline, SMTC), and NO donor (SNAP), were used to manipulate tissue NO. Baseline tissue NO was significantly higher in medulla (703+/-49 NM) than in cortex (231+/-17 nM). Minimal cortical and medullary NO current measured after maximal L-NAME dose (2.4 mg kg(-1) i.v.) was taken as tissue NO zero kevel. This dose decreased RBF and MBF significantly (-43%). SMTC, 1.2 mg kg(-1) h(-1) i.v., significantly decreased tissue NO by 105+/-32 nM in cortex and 546+/-64 nM in medulla, RBF and MBF decreased 30% and 20%, respectively. Renal artery infusion of SNAP, 0.24 mg kg(-1) min(-1) significantly increased tissue NO by 139+/-18 nM in cortex and 948+/-110 nM in medulla. Since inhibition of nNOS decreased medullary NO by 80% and MBF by 20% only, this isoform has probably minor role in the maintenance of medullary perfusion. [Abstract/Link to Full Text]

Jaworek J, Konturek SJ, Macko M, Kot M, Szklarczyk J, Leja-Szpak A, Nawrot-Porabka K, Stachura J, Tomaszewska R, Siwicki A, Pawlik WW
Endotoxemia in newborn rats attenuates acute pancreatitis at adult age.
J Physiol Pharmacol. 2007 Mar;58(1):131-47.
Bacterial endotoxin (lipopolysaccharide, LPS), at high concentration is responsible for sepsis, and neonatal mortality, however low concentration of LPS protected the pancreas against acute damage. The aim of this study was to investigate the effect of exposition of suckling rats to LPS on the course of acute pancreatitis at adult age. Suckling rat (30-40g) received intraperitoneal (i.p.) injection of saline (control) or LPS from Escherichia coli or Salmonella typhi (5, 10 or 15 mg/kg-day) during 5 consecutive days. Two months later these rats have been subjected to i.p. cearulein infusion (25 microg/kg) to produce caerulein-induced pancreatitis (CIP). The following parameters were tested: pancreatic weight and morphology, plasma amylase and lipase activities, interleukin 1beta (IL-1 beta), interleukin 6 (IL-6), and interleukin 10 (IL-10) plasma concentrations. Pancreatic concentration of superoxide dismutase (SOD) and lipid peroxidation products; malondialdehyde (MDA) and 4-hydroxynonenal (4-HNE) have been also measured. Caerulein infusion produced CIP in all animals tested, that was confirmed by histological examination. In the rats, which have been subjected in the neonatal period of life to LPS at doses 10 or 15 mg/kg-day x 5 days, all manifestations of CIP have been reduced. In these animals acute inflammatory infiltration of pancreatic tissue and pancreatic cell vacuolization have been significantly diminished. Also pancreatic weight, plasma lipase and alpha-amylase activities, as well as plasma concentrations of IL-1beta and IL-6 have been markedly decreased, whereas plasma anti-inflammatory IL-10 concentration was significantly increased in these animals as compared to the control rats, subjected in the infancy to saline injection instead of LPS. Caerulein-induced fall in pancreatic SOD concentration was reversed and accompanied by significant reduction of MDA + 4 HNE in the pancreatic tissue. The effects of LPS derived from E. coli or S. typhi were similar. Pretreatment of suckling rats with LPS at dose of 10 mg/kg-day x 5 days resulted in the most prominent attenuation of acute pancreatitis at adult age, whereas LPS at dose of 5 mg/kg-day x 5 days given to the neonatal rats failed to affect significantly acute pancreatitis induced in these animals 2 months later. We conclude that: 1/ Prolonged exposition of suckling rats to bacterial endotoxin attenuated acute pancreatitis induced in these animals at adult age. 2/ This effect could be related to the increased concentration of antioxidative enzyme SO in the pancreatic tissue and to the modulation of cytokines production in these animals. [Abstract/Link to Full Text]

Slomiany BL, Slomiany A
Interference by leptin with Helicobacter pylori lipopolysaccharide-induced cytosolic phospholipase A2 activation in gastric mucosal cells.
J Physiol Pharmacol. 2007 Mar;58(1):117-30.
Activation of cytosolic phospholipase A(2) (cPLA(2)) by bacterial LPS for the rapid release of arachidonic acid from membrane phospholipids is considered a key step in the generation of platelet-activating factor (PAF), recognized as the most proximal mediator of inflammatory events triggered by bacterial infection. In this study, we report on the role of leptin in modulation of the detrimental consequences of H. pylori LPS-induced cPLA(2) activation that result in the disturbances in gastric mucin synthesis. Employing gastric mucosal cells labeled with [(3)H] arachidonic acid, we show that H. pylori LPS-induced cPLA(2) activation, associated with up-regulation in apoptosis and PAF generation, and the impairment in gastric mucin synthesis, was subject to a dose-dependent suppression by leptin, as well as the inhibition by MAFP, a specific inhibitor of cPLA(2). A potentiation in the countering capacity of leptin on the LPS-induced up-regulation in apoptosis, arachidonic acid release and PAF generation was attained in the presence of ERK inhibitor, PD98059, while PI3K inhibitor, wortmannin had no effect. On the other hand, the prevention by leptin of the LPS detrimental effect on mucin synthesis was subject to suppression by wortmannin, an inhibitor of PI3K as well as the inhibitor of ERK, PD98059. Moreover, potentiation in the effect of leptin on the LPS-induced decrease in mucin synthesis was attained with cPLA(2) inhibitor, MAFP as well as PAF receptor antagonist, BN52020. The results of our findings point to H. pylori LPS-induced ERK-dependent cPLA(2) activation as a critical factor influencing the level of PAF generation, and hence the extent of pathological consequences of H. pylori infection on the synthesis of gastric mucin. Furthermore, we show that leptin counters the pathological consequences of H. pylori-induced cPLA(2) activation on gastric mucin synthesis through the involvement in signaling events controlled by MAPK/ERK and PI3K pathways. [Abstract/Link to Full Text]

Yildirim A, Sahin YN, Suleyman H, Yilmaz A, Yildirim S
The role of prednisolone and epinephrine on gastric tissue and erythrocyte antioxidant status in adrenalectomized rats.
J Physiol Pharmacol. 2007 Mar;58(1):105-16.
It has been believed that overproduction of free radicals and/or deficiency of antioxidant systems, and stress hormones may play a role in etiopathogenesis of many diseases, including gastric ulcer. This study evaluated whether there was an effect of adrenalectomy on lipid peroxidation [malondialdehyde (MDA)] and antioxidant [superoxide dismutase (SOD), glutathione peroxidase (GPX) and glutathione (GSH) levels] systems in gastric tissue and erythrocyte in rats. As well, the impacts of administration of prednisolone and epinephrine on these systems in adrenalectomized rats were investigated. Thirty-three rats were randomly grouped as sham-operated (group I), adrenalectomized (group II), adrenalectomized + prednisolone (group III) and adrenalectomized + epinephrine (group IV). After experimental procedures, blood and gastric tissues samples were taken from each animal in all groups. Colorimetric assays were employed to determine gastric tissue and erythrocyte levels of MDA and GSH, and SOD and GPX activities. Adrenalectomy in group II rats caused a marked decrease of SOD and GPX activities and MDA levels, and an increase of GSH levels in gastric tissue and erythrocyte, when compared to sham-operated rats. However, especially epinephrine injection after adrenalectomy resulted in a significantly increase of measured antioxidant enzyme activities and GSH levels in both gastric tissue and erythrocyte. These results indicate that adrenalectomy appeared to alter the levels of antioxidants and lipid peroxidation product in gastric tissue and erythrocyte. Thus, the present study provides a physiological regulatory role of adrenal gland in the maintenance of oxidant/antioxidant balance in gastric tissue and erythrocyte. [Abstract/Link to Full Text]

Mlynarik M, Makatsori A, Dicko I, Hinghofer-Szalkay HG, Jezova D
Postural changes associated with public speech tests lead to mild and selective activation of stress hormone release.
J Physiol Pharmacol. 2007 Mar;58(1):95-103.
We tested whether simulation of postural changes, which occur during public speech test procedures, activates cardiovascular system and stress hormone release that could interfere with the effect of psychosocial stress load. Young healthy male volunteers (n=8) underwent procedure imitating exactly all postural changes present in the psychosocial stress model based on public speech used in this laboratory (namely changes from sitting to standing and repeated sitting). Postural changes were associated with increases in heart rate, blood pressure, plasma concentrations of noradrenaline and aldosterone and elevation in plasma renin activity. In contrast to cardiovascular parameters, adrenocorticotropic hormone, cortisol and adrenaline, the main characteristics of hormonal response during mental stress, were not significantly influenced. The overall magnitude of all observed alterations was much smaller than that seen following mental stress procedures in our previous studies. This study provides evidence that changes in body posture during public speech test procedure influence hemodynamics and endocrine responses in a mild manner. Though this influence may represent a source of unspecific variance, substantial confounding effects on responses to the psychosocial component of the procedure are unlikely. In any case, models combining mental stressors and changes in body posture must be interpreted as complex stress stimuli. [Abstract/Link to Full Text]

Klenerová V, Sída P, Krejcí I, Hlinák Z, Hynie S
Effects of two types of restraint stress on spontaneous behavior of Sprague-Dawley and Lewis rats.
J Physiol Pharmacol. 2007 Mar;58(1):83-94.
The purpose of this study was to evaluate the action of two types of stressors in Sprague-Dawley (S-D) and Lewis (LEW) rats differing in their hypothalamic-pituitary-adrenal axis activity on locomotion and rearing in an open space. Exposure to restraint immobilization alone (IMO) or this immobilization combined with cold water (22 degrees C) immersion (IMO+C) lasted for 1 h and started 2 or 5 h before the test. To evaluate the acute and persisting effects of both stressors, four trials were performed in one-week intervals; rats were exposed to the stressors in trial 1 and 3. While in LEW rats both acute stressors produced reduction of locomotion and rearing in all stressed groups, S-D rats responded with a decrease of both parameters only after IMO+C presented 2 h before testing. Neither strain displayed a changed performance one week after the first stress exposure. One week after the second stress exposure rats of both strains exhibited a tendency to an increase of both parameters reaching the significance in several experimental groups. The findings indicate: a) the IMO+C produced stronger behavioral alteration than IMO alone; b) the behavioral responses to stressors were more pronounced in LEW compared to S-D strain; c) change from the reduction of activity to its increase may be interpreted as bi-directional manifestation of long-term effects of immobilization stress. [Abstract/Link to Full Text]

Chabowski A, Zmijewska M, Górski J, Bonen A, Kami?ski K, Winnicka MM
Effect of IL-6 deficiency on myocardial expression of fatty acid transporters and intracellular lipid deposits.
J Physiol Pharmacol. 2007 Mar;58(1):73-82.
IL-6 is a biologically active substance and is thought to contribute to the development of obesity. Recent findings suggest that susceptibility to intracellular lipid accumulation is to a large extent determined by changes in the expression of fatty acid transporters such as FAT/CD36, FABPpm and FATP-1. The aim of the present study was to determine the effect of IL-6 deficiency on the expression of fatty acid transporters, as well as, assess the concomitant changes in intracellular lipids. We found that Il-6 deficiency upregulated the myocardial expression of FAT/CD36 (+40%) and did not significantly affect the content of FABPpm and FATP-1 (+15% and +5% respectively). Although no change in the intramyocardial total lipid content was noted, there was a significant increase in the intracellular content of both free fatty acid (FFA), diacylglicerol (DG) and ceramide fractions (+45%, +37% and +48%, respectively) in hearts from IL-6 -/- mice. A trend for IL-6 deficiency to increase in saturated FA species in these fractions was also observed (+8%, +12% and +10%, respectively). In contrast, IL-6 deficiency has no effect on the content of monounsaturated fatty acid (MUFA) and polyunsaturated fatty acid (PUFA) species in each intramyocardial lipid fractions examined. These findings suggest that IL-6 deficiency results in 1) upregulation of myocardial content of FAT/CD36, 2) the increase in the content of biologically active lipid pools (FFA, DG and ceramide). This lipid accumulation with concomitant trend for increase in the saturation status of these lipid fractions may, at least in part, provide a factor related to the development of intramyocardial lipotoxicity, observed in obese individuals. [Abstract/Link to Full Text]

Baranowski M, B?achnio A, Zabielski P, Górski J
PPARalpha agonist induces the accumulation of ceramide in the heart of rats fed high-fat diet.
J Physiol Pharmacol. 2007 Mar;58(1):57-72.
It was shown that high-fat feeding of mice with cardiac-specific overexpression of peroxisome proliferator-activated receptor (PPAR) alpha but not wild type animals leads to the accumulation of ceramide (an important mediator of lipotoxicity) in the heart [Finck et al. 2003 Proc Natl Acad Sci USA]. To investigate the mechanism of this phenomenon we examined the effects of PPARalpha activation on ceramide metabolism in the myocardium. Male Wistar rats were fed either a standard chow or a high-fat diet. Each group was divided into two subgroups: control and treated with selective PPARalpha activator - WY-14643. In the rats fed on the standard diet WY-14643 did not affect the myocardial content of sphingomyelin and ceramide but reduced the content of sphinganine and sphingosine. It also inhibited the activity of neutral sphingomyelinase and increased the activity of acid sphingomyelinase, whereas the activity of ceramidases and serine palmitoyltransferase (SPT) remained stable. High-fat diet itself did not affect the content of the examined sphingolipids. However, it reduced the activity of sphingomyelinases and ceramidases having no effect on the activity of SPT. Administration of WY-14643 to this group significantly increased the content of myocardial free palmitate, ceramide, sphingomyelin and the activity of SPT. Our results demonstrated that PPARalpha activation modulates myocardial ceramide metabolism and leads to the accumulation of ceramide in the heart of the high-fat fed rats due to its increased synthesis de novo. [Abstract/Link to Full Text]

Recent Articles in Polish Journal of Pharmacology and Pharmacy

Czapski GA, Caka?a M, Kopczuk D, Kami?ska M, Strosznajder JB
Inhibition of nitric oxide synthase prevents energy failure and oxidative damage evoked in the brain by lipopolysaccharide.
Pol J Pharmacol. 2004 Sep-Oct;56(5):643-6.
The inducible nitric oxide synthase (iNOS) plays an important role in endotoxic shock. However,little is known about the involvment of constitutive isoform(s) of NOS (cNOS). The aim of this study was to determine the role of cNOS in the mouse brain after lipopolysaccharide (LPS) injection. Concentrations of nicotinamide adenine dinucleotide (NAD(+)), carbonyl group and thiobarbituric acid reactive substances were determined spectrophotometrically, cNOS mRNA was evaluated by RT-PCR. Our data showed that LPS significantly decreased NAD(+) level, and enhanced protein and lipid oxidation, but had no effect on cNOS mRNA expression. Inhibitors of cNOS protected the cells against alterations evoked by LPS, suggesting involvement of cNOS isoforms in pathology. [Abstract/Link to Full Text]

Wesierska-Gadek J, Gueorguieva M, Wojciechowski J, Horky M
Cell cycle arrest induced in human breast cancer cells by cyclin-dependent kinase inhibitors: a comparison of the effects exerted by roscovitine and olomoucine.
Pol J Pharmacol. 2004 Sep-Oct;56(5):635-41.
Cyclin-dependent kinases (CDKs) are serine/threonine kinases that play a key role in the regulation of the cell cycle progression. In proliferating cells, distinct CDKs activated upon complexing with specific cyclins and upon site-specific phosphorylation coordinate in an orchestrated way the appropriate transition between consecutive phases of the cell cycle. Aberrant expression or altered activity of distinct CDK complexes results in escape of cells from the cell cycle control and leads to malignant transformation. Therefore, the inhibition of CDKs in malignant cells provides a new strategy in the fight against cancer. Recently, selective CDK inhibitors targeting distinct CDKs were developed. They represent promising anti-cancer drugs due to their strong anti-proliferative efficacy combined with a relative low direct cytotoxicity. The aim of this study was to compare the effect of two related CDK inhibitors: roscovitine (ROSC) and olomoucine (OLO) on the cell cycle progression in human breast cancer MCF-7 cells. Both examined CDK inhibitors differentially affected the cell cycle progression in MCF-7 cels. Whereas ROSC arrested cells in G(2)/M, OLO inhibited cells at S to G(2) transition and increased the number of cells residing in the S-phase. Moreover, both CDK inhibitors modulated the cell cycle progression with distinct kinetics. Accumulation of G(2)/M-arrested cells beginning 6 h after exposure of cells to ROSC coincided with a strong up-regulation of the p53. Interestingly, ROSC triggered apoptosis in MCF-7 cells by activation of mitochondrial pathway. Loss of the integrity of mitochondrial membrane observed after exposure of cells to ROSC for 6 h led to release of distinct mitochondrial proteins, e.g. apoptosis inducing factor (AIF). In contrast to ROSC, OLO-induced cell cycle changes could be detected after 12 h of the treatment. OLO did not up-regulate p53 protein. It indicates that both examined CDK inhibitors are selective and block the cell cycle progression of human breast carcinoma cells at different phases. [Abstract/Link to Full Text]

Keil U, Bonert A, Marques CA, Strosznajder JB, Müller-Spahn F, Müller WE, Eckert A
Elevated nitric oxide production mediates beta-amyloid-induced mitochondria failure.
Pol J Pharmacol. 2004 Sep-Oct;56(5):631-4.
Mitochondrial dysfunction has been identified in a large proportion of neurodegenerative disorders including Alzheimer's disease (AD). In addition, the involvement of nitric oxide (NO) has been implicated in the pathogenesis of AD. Thus, we investigated the effects of the Swedish double mutation (K670M/N671L) in the beta-amyloid precursor protein (APPsw) on NO levels and mitochondrial function in PC12 cells. Interestingly, APPsw PC12 cells showed increased NO levels, decreased cytochrome C oxidase activity and reduced ATP levels compared to wild-type APP bearing cells and empty vector transfected cells. On the basis of our data, we propose a hypothetical sequence of events linking amyloid beta-peptide and NO production with mitochondria failure. [Abstract/Link to Full Text]

de Melo GA, Sartoretto JL, Caparroz-Assef SM, Bersani-Amado CA, Fortes ZB, Cuman RK
Participation of endogenous corticosteroids in inflammatory response in type 2 streptozotocin diabetic rats.
Pol J Pharmacol. 2004 Sep-Oct;56(5):617-9.
The inflammatory response is decreased in diabetic animals. After adrenals removal this impaired response in type 2 diabetic rats evaluated by pleurisy and vascular permeability tests was restored. Our studies demonstrate that endogenous corticosteroids play a partial role in the impaired inflammatory response in type 2 streptozotocin diabetic rats. [Abstract/Link to Full Text]

Szczepanik AM, Wordliczek J, Serednicki W, Siedlar M, Czupryna A
Pentoxifylline does not affect nociception if administered postoperatively.
Pol J Pharmacol. 2004 Sep-Oct;56(5):611-6.
Proinflammatory cytokines, such as tumor necrosis factor alpha (TNF-alpha), interleukin-1 (IL-1) and interleukin-6 (IL-6), act as mediators of post-injury inflammation and increase pain sensitivity. Pentoxifylline (PTX) has the property of inhibiting TNF-alpha, IL-1, and IL-6 production. Previous studies revealed that the pre-injury or preoperative administration of PTX inhibited consequent hyperalgesia or postoperative pain. The aim of the study was to determine, if postoperative PTX administration affects postoperative pain. A group of 40 patients undergoing laparotomic cholecystectomy received postoperatively PTX at 10 mg/kg or placebo directly after the termination of general anesthesia. There were no differences in postoperative pain, analgesic drug requirement or TNF-alpha and IL-6 serum levels between the groups. [Abstract/Link to Full Text]

Bujalska M
Effect of nitric oxide synthase inhibition on antinociceptive action of different doses of acetaminophen.
Pol J Pharmacol. 2004 Sep-Oct;56(5):605-10.
As demonstrated in previous studies, both cyclooxygenases (COXs) and nitric oxide synthases (NOS) localized peripherally and/or centrally participate in the antinociceptive action of acetaminophen (ACETA). We showed that opioidergic system(s) was involved in the mechanism of ACETA action, as well. In previous and recent studies, the changes in nociceptive threshold were estimated using a mechanical and chemical stimulus. In this study, the influence of nonspecific inhibitor of NOS [N(G)-nitro-L-arginine (L-NOArg)] on antinociceptive action of ACETA administered icv and it was studied in rats. ACETA increased threshold for electrical stimuli, however, its analgesic activity was not dose-dependent. Independently of the route of administration, the existence of a ceiling dose of ACETA was observed above which the activity of ACETA was self-limited. Pretreatment with L-NOArg (ip) markedly increased the action of higher doses of ACETA. It suggests that the attenuation of analgesic action of higher doses of ACETA may be due to increased activity of NOS. [Abstract/Link to Full Text]

Lukivskaya O, Lis R, Zwierz K, Buko V
Effect of the nitric oxide donor and the nitric oxide synthase inhibitor on the liver of rats with chronic hepatitis induced by dimethylnitrosamine.
Pol J Pharmacol. 2004 Sep-Oct;56(5):599-604.
The present study was designed to examine the effects of the donor of nitric oxide (NO), NaNO(2) and the inhibitor of NO synthase, N(omega)-nitro-L-arginine (L-NNA), on the development of dimethylnitrosamine (DMNA)-induced chronic hepatitis in rats. L-NNA decreased rat survival and enhanced the severity of hepatic encephalopathy in the DMNA-treated animals. The aggravation of the morphological signs of hepatitis, the activation of serum alanine aminotransferase and cytosolic superoxide dismutase activities and the increase in the liver malondialdehyde content were observed in this group. The treatment with NaNO(2) improved liver morphology, decreased serum marker enzyme activities, lowered the activities of alpha-D-mannosidase and N-acetyl-beta-D-glucosaminidase compared to the DMNA-treated group. The results of the morphological and biochemical studies suggest that L-NNA increased DMNA-induced liver damage, whereas NaNO(2) partially prevented the development of chronic hepatitis. It is proposed that the opposite effects of L-NNA and NaNO(2) are partially explained by a modulation of the free radical-dependent processes in the liver. [Abstract/Link to Full Text]

Zadrozniak M, Sekowski A, Czuczwar SJ, Borowicz KK
Effect of MPEP, a selective mGluR5 antagonist, on the antielectroshock activity of conventional antiepileptic drugs.
Pol J Pharmacol. 2004 Sep-Oct;56(5):595-8.
MPEP, a selective non-competitive antagonist of group I metabotropic glutamate receptor subtype 5 (mGluR5), administered at doses ranging from 0.75 to 1 mg/kg, failed to influence the electroconvulsive threshold in mice. However, when administered at higher doses (1.25 and 1.5 mg/kg), it significantly increased the threshold. Moreover, MPEP (applied at its highest subprotective dose of 1 mg/kg) did not affect the protective action of valproate, carbamazepine, diphenylhydantoin and phenobarbital against maximal electroshock-induced seizures in mice. The presented results indicate that mGluR5 antagonists should not be considered as good candidates for add-on therapy of generalized seizures. [Abstract/Link to Full Text]

Prakasam A, Sethupathy S, Pugalendi KV
Influence of Casearia esculenta root extract on protein metabolism and marker enzymes in streptozotocin-induced diabetic rats.
Pol J Pharmacol. 2004 Sep-Oct;56(5):587-93.
The present study investigated the possible protective effects of Casearia esculenta root extract on certain biochemical markers in streptozotocin (STZ)-induced diabetes in rats. STZ treatment (50 mg/kg, ip) caused a hyperglycemic state, that led to various physiological and biochemical alterations. Blood levels of glucose, urea, uric acid and creatinine, plasma levels of albumin and albumin/globulin ratio and the activities of diagnostic marker enzymes aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP) and gamma-glutamyltranspeptidase (gamma-GT) in plasma, liver and kidney were markedly altered in STZ diabetic rats. Oral administration of C. esculenta (200 and 300 mg/kg) for 45 days restored all these biochemical parameters to near normal levels. Thus, the present results have shown that C. esculenta root extract has the antihyperglycemic effect and consequently may alleviate liver and renal damage associated with STZ-induced diabetes in rats. [Abstract/Link to Full Text]

Kalpana C, Menon VP
Modulatory effects of curcumin on lipid peroxidation and antioxidant status during nicotine-induced toxicity.
Pol J Pharmacol. 2004 Sep-Oct;56(5):581-6.
Nicotine, a pharmacologically active substance in tobacco, has been identified as a major risk factor for lung diseases. In the present study, we evaluated the protective effects of curcumin on tissue lipid peroxidation and antioxidants in nicotine-treated Wistar rats. Lung toxicity was induced by subcutaneous injection of nicotine at a dose of 2.5 mg/kg (5 days a week, for 22 weeks). Curcumin (80 mg/kg) was given simultaneously by intragastric intubation for 22 weeks. The enhanced level of tissue lipid peroxides in nicotine-treated rats was accompanied by a significant decrease in the levels of ascorbic acid, vitamin E, reduced glutathione, glutathione peroxidase, superoxide dismutase and catalase. Administration of curcumin significantly lowered the level of lipid peroxidation and enhanced the antioxidant status. The results of the present study suggest that curcumin exerts its protective effect against nicotine-induced lung toxicity by modulating the extent of lipid peroxidation and augmenting antioxidant defense system. [Abstract/Link to Full Text]

Pytlik M, Kaczmarczyk-Sedlak I, Sliwi?ski L, Janiec W, Rymkiewicz I
Effect of concurrent administration of alendronate sodium and retinol on development of changes in histomorphometric parameters of bones induced by ovariectomy in rats.
Pol J Pharmacol. 2004 Sep-Oct;56(5):571-9.
Retinol is a commonly used vitamin, especially by elderly people. Alendronate sodium, an aminobisphosphonate, is a potent antiresorptive drug used in the treatment of osteoporosis in postmenopausal women. Frequently, alendronate sodium and retinol are used concurrently. There are no reports on the interaction between alendronate sodium and retinol. The aim of the present study was to investigate the effect of concurrent administration of alendronate sodium and retinol on bone remodeling in ovariectomized rats. The histomorphometric parameters of long bones were studied. The experiments were carried out on 3-month-old Wistar rats, divided into 7 groups: I (C) - sham operated control rats, II (OVX) - ovariectomized control rats, III (OVX + ALN) - ovariectomized rats + alendronate sodium (3 mg/kg po), IV (OVX + R-1) - ovariectomized rats + retinol (700 IU/kg po), V (OVX + R-2) - ovariectomized rats + retinol (3500 IU/kg po), VI (OVX + ALN + R-1) - ovariectomized rats + alendronate sodium (3 mg/kg po) + retinol (700 IU/kg po), VII (OVX + ALN + R-2) - ovariectomized rats + alendronate sodium (3 mg/kg po) + retinol (3500 IU/kg po). The drugs were administered to the rats daily by oral gavage (alendronate sodium in the morning, retinol in the afternoon) for 28 days. Body mass gain, bone mass, mineral content in the tibia, femur and L-4 vertebra, histomorphometric parameters of the right tibia (width of osteoid, periosteal and endosteal transverse growth, area of the transverse cross section of the bone marrow cavity and the cortical bone) and the right femur (width of epiphyseal and metaphyseal trabeculae, width of epiphyseal cartilage) were studied. Bilateral ovariectomy induced osteopenic skeletal changes in mature female rats. Alendronate sodium administered at a dose of 3 mg/kg po daily inhibited the development of changes induced by ovariectomy in the skeletal system of rats. Retinol, especially administered at the dose of 3500 IU/kg daily, intensified the changes in the osseous system caused by estrogen deficiency in rats. Retinol administered concurrently with alendronate sodium attenuated the antiresorptive effect of alendronate sodium on the skeletal system in ovariectomized rats. [Abstract/Link to Full Text]

Kowalski J, Labuzek K, Herman ZS
Flupentixol and trifluperidol reduce interleukin-1 beta and interleukin-2 release by rat mixed glial and microglial cell cultures.
Pol J Pharmacol. 2004 Sep-Oct;56(5):563-70.
Neuroleptics penetrate into the brain, where they act not only on neurons but probably also on glial cells. In the available literature, there are no reports on the effect of neuroleptics on cytokine release in glia cultures. The aim of this study was to evaluate the effect of neuroleptics on the release of proinflammatory cytokines (IL-1beta and IL-2) by mixed glial and microglial cell cultures. Trifluperidol at 20 and 2 muM reduced IL-beta secretion by mixed glial cultures after 3 days of exposure. Trifluperidol at 20, 2 and 0.2 muM diminished IL-beta secretion after 1 day of incubation. Trifluperidol at 20 and 2 muM reduced IL-2 release after 1 and 3 days of exposure. Flupentixol at 20 and 2 muM reduced IL-1beta by mixed glial cell cultures after 3 days of exposure. Flupentixol at 20, 2 and 0.2 muM caused diminution of IL-1beta release after 1 day of exposure. Flupentixol at 20 and 2 muM reduced IL-2 release after 1 day of incubation. Flupentixol at 20, 2 and 0.2 muM diminished IL-2 release after 3 days of exposure. Flupentixol at 20, 10, 2 and 0.2 muM reduced IL-1beta release by microgial cell cultures. Flupentixol at 20, 10 and 2 muM reduced release of IL-2 by microglial cells after 1 day of exposure. The results of the present study suggest that neuroleptics have an inhibiting effect on the release of glial cytokines, but clinical significance this results remains to be elucidated. [Abstract/Link to Full Text]

Kubera M, Budziszewska B, Jaworska-Feil L, Basta-Kaim A, Le?kiewicz M, Tetich M, Maes M, Kenis G, Marciniak A, Czuczwar SJ, Jag?a G, Nowak W, Laso? W
Effect of topiramate on the kainate-induced status epilepticus, lipid peroxidation and immunoreactivity of rats.
Pol J Pharmacol. 2004 Sep-Oct;56(5):553-61.
Topiramate, a new anticonvulsant, has been reported to possess neuroprotective effects in both in vivo and in vitro experiments. In the present study, the effect of topiramate (40 and 80 mg/kg ip) on the fully developed kainate-induced status epilepticus was evaluated in the rat. Injection of kainate (15 mg/kg ip) evoked recurrent limbic seizures which lasted several hours. Topiramate injected 1.5 h after kainate administration had no effect on the seizures and mortality of the animals. Biochemical study revealed that at 80 mg/kg ip, topiramate significantly attenuated the kainate-induced lipid peroxidation in the piriform cortex and showed similar tendency in the frontal cortex. Besides the central nervous system, the kainate-induced seizures evoked significant changes in immunoreactivity, such as reduction in thymus weight and the proliferative activity of splenocytes, and the splenocyte-increased production of interleukin-10, but not interferon-gamma. Topiramate did not affect the kainate-induced reduction in thymus weight, but attenuated changes in the proliferative activity of splenocytes. It is concluded that topiramate, when given during the fully developed kainate-induced status epilepticus in rats, has no effect on seizures, but attenuates lipid peroxidation in piriform cortex and prevents certain changes in immunoactivity. [Abstract/Link to Full Text]

Wnek W, Zajaczkowska R, Wordliczek J, Dobrogowski J, Korbut R
Influence of pre-operative ketoprofen administration (preemptive analgesia) on analgesic requirement and the level of prostaglandins in the early postoperative period.
Pol J Pharmacol. 2004 Sep-Oct;56(5):547-52.
The aim of this study was to examine the effect of ketoprofen used in preemptive analgesia on the intensity of pain and requirement for analgesics in the perioperative period. Sixty patients scheduled for elective lumbar disc prolapse surgery were randomly divided into two groups. In the PRE group (n = 30) ketoprofen was administered one hour before incision. In the POST group ( n = 30) ketoprofen was used immediately after the surgery. The operation was performed under general anesthesia. Postoperative analgesia was realized by NCA (Nurse Controlled Analgesia) and the "required" dose of ketoprofen was 100 mg. After the operation, pain intensity was measured using visual-analog scale (VAS), ketoprofen requirements, the time to the first dose of ketoprofen, and levels of prostaglandin E(2) (PEG(2)) in blood serum were compared. There were no differences between the groups in the VAS pain scores, and levels of PGE(2) in blood serum. However, in patients of PRE group who had received preemptive analgesia, a significantly lower total consumption of ketoprofen, as compared with POST group, was observed between 12th and 36th postoperative hours. It was also found that the time which elapsed between the end of the operation and the first NCA activation was significantly shorter in the PRE group, as compared with the POST group. The results of our study confirm the possibility of modifying the nociception process in the perioperative period through preemptive analgesia by ketoprofen. [Abstract/Link to Full Text]

Bujalska M
Effect of nonselective and selective opioid receptors antagonists on antinociceptive action of acetaminophen [part III].
Pol J Pharmacol. 2004 Sep-Oct;56(5):539-45.
The influence of naloxone (NAL), a competitive antagonist of mu, kappa, delta and sigma receptors; D-Phe-Cys-Tyr-D-Trp-Orn-Thr-Pen-Thr-NH(2) (CTOP), selective antagonist of mu-opioid receptors; nor-binaltorphimine (NOR-BNI), a potent and higly selective kappa opioid receptor antagonist; naltrindole (NTI), a delta-opioid receptor antagonist and naltriben (NTB), a highly selective delta(2)-opioid receptor antagonist on antinociceptive action of acetaminophen (ACETA) was studied in rats. NAL administered intraperitoneally (ip) or intracerebroventricularly (icv), and CTOP and NOR-BNI administered icv, markedly decreased the antinociceptive activity of the high dose of ACETA (400 mg/kg). Pretreatment with NTI (sc), as well as with naloxone (it), and NTB (it) slightly but significantly attenuated the ACETA antinociception. The possible involvement of the opioidergic systems in antinociceptive activity of ACETA is discussed. [Abstract/Link to Full Text]

Car H, Wi?niewska RJ, Wi?niewski K
2R,4R-APDC influence on hypoxia-induced impairment of learning and memory processes in passive avoidance test.
Pol J Pharmacol. 2004 Sep-Oct;56(5):527-37.
We investigated the effects of 2R,4R-APDC, a selective group II metabotropic glutamate receptor (II mGluR) agonist, on certain behaviors in rats subjected and non-subjected to hypoxia. Short-term hypoxia was used as a model of experimentally induced amnesia. 2R,4R-APDC given intracerebroventricularly (icv) at doses of 1 mumol and 100 nmol decreased the number of crossings and rearings in the open field, impaired acquisition and consolidation but improved retrieval in the passive avoidance tests. It also shortened the time spent in open arms and prolonged the time spent in closed arms, reduced the number of open and closed arms entries in an elevated "plus" maze, which is a measure of anxiety. Four-minute hypoxia (2% O(2), 98% N(2)) retrieval of conditioned responses, and exhibited an anxiogenic effect in the elevated "plus" maze in rats, i.e. it reduced the time spent in open arms and the number of entries to closed and open arms. 2R,4R-APDC effect on locomotor and exploratory activity was not changed after hypoxia, i.e. we observed inhibition of motility. This agonist of II mGluRs used at both doses before hypoxia significantly improved acquisition and retrieval, and had dual effect on consolidation, viz. at a dose of 1 mumol, it impaired this process and at a dose of 100 nmol it improved it. In the elevated "plus" maze, rats pretreated with 2R,4R-APDC and then subjected to hypoxia shortened the time spent in open arms and prolonged the time spent in closed arms, reduced the time spent in open arms, i.e. the drug exhibited anxiogenic effect. We conclude, therefore, that 2R,4R-APDC itself impaired acquisition and consolidation, enhanced retrieval but in rats undergoing hypoxia, it improved acquisition, retrieval and when used at the dose of 100 nmol enhanced consolidation. 2R,4R-APDC had beneficial effect in hypoxia-induced memory impairment in passive avoidance test. [Abstract/Link to Full Text]

Rogóz Z, Skuza G, K?lodzi?ska A
Anxiolytic- and antidepressant-like effects of 7-OH-DPAT, preferential dopamine D3 receptor agonist, in rats.
Pol J Pharmacol. 2004 Sep-Oct;56(5):519-26.
The aim of the present study was to examine a potential anxiolytic- and antidepressant-like action of (+)-7-hydroxy-2-(di-n-propylamino)tetralin hydrobromide (7-OH-DPAT), a preferential dopamine D(3) receptor agonist, and N-{4-[4-(2-methoxyphenyl)-1-piperazinyl]butyl}-2-naphthylcarboxamide (BP 897), a partial dopamine D(3) receptor agonist, in male Wistar rats. Diazepam or imipramine were used as reference compounds. The anxiolytic-like effect of those drugs was tested in the elevated plus-maze test. The antidepressant-like effect was estimated using the forced swimming test. The obtained results showed that 7-OH-DPAT at low doses and BP 897 (like diazepam) induced anxiolytic-like effects in the elevated plus-maze test. 7-OH-DPAT, BP 897 and diazepam, tested at the doses effective in the model of anxiolytic-like actions, did not affect motor coordination. Moreover, 7-OH-DPAT at higher doses, like imipramine, showed antidepressant-like effect, significantly reducing immobility time in the forced swimming test. Combined treatment with 7-OH-DPAT and imipramine induced a stronger effect in Porsolt's test than administration of either drug alone, but did not increase the locomotor activity. In contrast, BP 897 was inactive in the forced swimming test when given alone but it potentiated the antidepressant-like effect of imipramine. These data suggest that preferential D(3) receptor agonists may play a role in the therapy of anxiety and/or depression, however, further studies are necessary to elucidate the mechanism of these actions. [Abstract/Link to Full Text]

Bia?ek M, Zaremba P, Borowicz KK, Czuczwar SJ
Neuroprotective role of testosterone in the nervous system.
Pol J Pharmacol. 2004 Sep-Oct;56(5):509-18.
Testosterone -- the gonadal sex steroid hormone plays an important role in the central nervous system (CNS) development. One of the less known testosterone actions is neuroprotection. There are some evidences supporting the hypothesis that testosterone may act protectively in neurodegenerative disorders, e.g. Alzheimer's disease (AD), mild cognitive impairment (MCI) or depression. Androgens alter also the morphology, survival and axonal regeneration of motor neurons. These hormones accelerate the regeneration of hamster facial nerve and anterior tibialis sciatic nerve in rabbits following crush axotomy. Androgens exert trophic action in laryngeal motor nucleus of Xenopus laevis. Testosterone is linked to an increase in neuron somal size, neuritic growth, plasticity and synaptogenesis in both motoneurons of the spinal nucleus of the bulbocavernosus and several populations of pelvic autonomic neurons. The hormone reduced the extent of spinal cord damage in vitro. There are also evidences against the neuroprotective action of testosterone. Testosterone does not protect against methamphetamine-induced neurotoxicity of the dopaminergic system in mice and does not provide significant neuroprotection against glutamate-induced neurotoxicity. Androgens do not prevent striatal dopamine depletion induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) in mice. Although the role of testosterone in the CNS is still poorly understood, accumulating evidence suggests that testosterone may create a future treatment for MCI and related cognitive diseases, including dementia and may influence motor neuron regeneration in adulthood. Androgen replacement therapy in selected male populations may hold therapeutic promise for the prevention and/or treatment of age-related disorders associated with neuronal injury. [Abstract/Link to Full Text]

Kulig K, Nowicki P, Malawska B
Influence of the absolute configuration on pharmacological activity of antihypertensive and antiarrhythmic drugs.
Pol J Pharmacol. 2004 Sep-Oct;56(5):499-508.
Chirality is a fundamental property of biological systems and reflects the underlying asymmetry of matter. Interactions of drugs with receptors, enzymes or binding sites have long been known to be stereoselective, and it is increasingly recognized that both pharmacodynamic and pharmacokinetic events contribute to the overall clinically observed stereoselectivity. The pharmacological activity may reside only in one enantiomer, while the second one may be inactive or have desirable or undesirable activity. Two isomers may be nearly identical both in qualitative and quantitative aspects of pharmacological activity. The activity of particular enantiomers may differ only at the quantitative level. It is also possible that a particular enantiomer displays qualitatively different mode of action than the second one. This review describes the influence of the absolute configuration on pharmacological activity of the selected currently used or being under investigation drugs acting on cardiovascular system, especially as the antihypertensive and antiarrhythmic agents. [Abstract/Link to Full Text]

Szewczuk-Bogus?awska M, Kiejna A, Besz?ej JA, Orzechowska-Juzwenko K, Milejski P
Doxepin inhibits CYP2D6 activity in vivo.
Pol J Pharmacol. 2004 Jul-Aug;56(4):491-4.
OBJECTIVE: Doxepin is a tricyclic antidepressant formulated as a mixture of E-(trans) and Z-(cis) stereoisomers. Cytochrome P450 2D6 (CYP2D6) catalyzes the hydroxylation of E-doxepin and E-N-desmethyldoxepin stereospecically. There is evidence that tricyclic antidepressants might inhibit CYP2D6 activity but there is no data about the influence of doxepin on CYP2D6. MATERIALS AND METHODS: Eleven patients diagnosed with depression according to ICD-10 criteria were included in the study. After wash-out period, before doxepin treatment, sparteine metabolic ratio (MR1) was assessed. After 2-weeks of doxepin treatment, MR2 was estimated. Sparteine and its metabolites were determined in urine by gas chromatographic method of Eichelbaum et al. RESULTS: Based on MR1 values, 10 patients were classified as EM (extensive metabolizers) and 1 patient as PM (poor metabolizer). During the study, after doxepin treatment, none of patients has changed phenotype status. However, MR2 values were statistically significantly higher than MR1. CONCLUSION: These results show the inhibitory effect of doxepin on CYP2D6 activity and may be of clinical value, especially in polymedicated patients treated with other CYP2D6 substrates or inhibitors. [Abstract/Link to Full Text]

Dmoszy?ska A, Podhorecka M, Roli?ski J, Soroka-Wojtaszko M
Influence of lovastatin on BCL-2 and BAX expression by plasma cells and T lymphocytes in short-term cultures of multiple myeloma bone marrow mononuclear cells.
Pol J Pharmacol. 2004 Jul-Aug;56(4):485-9.
Lovastatin (LOV), until now largely used for the treatment of hypercholesterolemia is a new promising drug in multiple myeloma (MM), however, the precise mechanism of its antitumor activity is not clear yet. It is probable that this effect is mediated by down-regulation of BCL-2 expression. In this study, we analyzed BCL-2 and BAX expression in cells of MM patients exposed to LOV in short-term culture. The obtained results indicate an increase in susceptibility to apoptosis both in CD138+ malignant cells and CD8+ T lymphocytes. Interestingly, such a tendency was confirmed in vivo in MM patient subjected to 3 cycles of LOV therapy. [Abstract/Link to Full Text]

Wyska E, Szymura-Oleksiak J, Opoka W, Ba? B, Niewiara E, Pomierny L, Dyba?a M, Nowak G
Pharmacokinetic interaction after joint administration of zinc and imipramine in forced swim test in mice.
Pol J Pharmacol. 2004 Jul-Aug;56(4):479-84.
Recent preclinical and clinical data indicate beneficial role of zinc in the antidepressant treatment. To evaluate the mechanism of interaction between zinc and antidepressants, in the present study we examined the brain zinc, imipramine and desipramine concentrations in mice treated with combinations of zinc and imipramine and subjected to the forced swim test. We have chosen doses of zinc (10 mg/kg) and imipramine (15 mg/kg) which we have previously found to be ineffective in the forced swim test when given alone. However, when administered jointly, a significant reduction in the immobility time in this test was demonstrated. In the present study, we demonstrated a significant ca. 60% reduction in the brain desipramine and non-significant reduction (ca. 40%) in brain imipramine concentrations in the group of animals treated with zinc plus imipramine compared with animals treated with imipramine alone. The brain zinc concentration in the zinc plus imipramine group was reduced when compared with the group treated with zinc or imipramine alone. Since there was no increase in brain imipramine/desipramine or zinc brain concentration after combined zinc and imipramine treatment, the data suggest that pharmacodynamic rather than pharmacokinetic interaction between zinc and imipramine is responsible for behavioral effect in the forced swim test. [Abstract/Link to Full Text]

Budzisz E, Krajewska U, Rózalski M
Cytotoxic and proapoptotic effects of new Pd(II) and Pt(II)-complexes with 3-ethanimidoyl-2-methoxy-2H-1,2-benzoxaphosphinin-4-ol-2-oxide.
Pol J Pharmacol. 2004 Jul-Aug;56(4):473-8.
The new palladium (II) and platinum (II) complexes 2 and 3 with 3-ethanimidoyl-2-methoxy-2H-1,2-benzoxaphospinin-4-ol-2-oxide 1 were synthesized and their physicochemical and pharmacological properties were determined. The cytotoxic effects of these complexes were examined on the two human leukemia cell lines, HL-60 and NALM-6. Pd-complex 2 and Pt-complex 3 exerted significant cytotoxic activity. The effects exhibited by these two complexes were comparable to those reported for cis-platin and carboplatin. On the other hand, ligand 1 was not cytotoxic. As determined by caspase-3 activity assay, compounds 2 and 3 differed slightly in their mode of induction of the programmed cell death. [Abstract/Link to Full Text]

Wójtowicz AK, Gregoraszczuk EL, Ptak A, Falandysz J
Effect of single and repeated in vitro exposure of ovarian follicles to o,p'-DDT and p,p'-DDT and their metabolites.
Pol J Pharmacol. 2004 Jul-Aug;56(4):465-72.
The aim of the presented study was to compare the effect of o,p'-DDT [1,1-dichloro-2,2-bis-(p,p'-chlorophenyl)-ethylene] and p,p'-DDT [1,1,1-trichloro-2,2-bis-(p-chlorophenyl)-ethane] and their metabolites DDE and DDD on estradiol secretion by ovarian follicles, the target organs of environmental estrogens. Theca interna (Tc) and granulosa cells (Gc) were collected from medium size porcine follicles and cultured as a monolayer. The cells were initially cultured for 24 h to allow attachment to the plates and then media were changed for the new ones and o,p'-DDT and p,p'-DDT and their metabolites: o,p'-DDE, p,p'-DDE and o,p'-DDD were added at doses of 4, 40, 400 ng and 4 microg/ml medium to investigate dose-dependent effects. Media were collected after 24 h and frozen for estradiol content determination. When the effect of single and repeated exposure was investigated, the lowest dose of 4 ng/ml and the highest one of 4 microg/ml were chosen on the basis of the results of Experiment 1. o,p'-DDT exerted antiestrogenic action at all doses used while its metabolites and p,p'-DDT and its metabolites decreased estradiol secretion only when present in the medium at a dose of 4 ng/ml. The highest doses caused the increase in estradiol secretion. Parent o,p'-DDT and its metabolites showed antiestrogenic action after single exposure to 4 ng/ml while parent p,p'-DDT and its metabolites caused estrogenic action. All investigated compounds, except o,p'-DDT, increased estradiol secretion after single exposure to the dose of 4 microg/ml. Repeated exposure resulted in a massive antiestrogenic action of all investigated chemicals. In conclusion, our study points to time-dependent effect of DDT and its metabolites on ovarian follicles with the strongest estrogenic properties observed after single exposure and antiestrogenic action caused by repeated exposure. Given the duration of folliculogenesis, one can imagine many different potential mechanisms by which DDT could influence steroidogenesis. [Abstract/Link to Full Text]

Borowicz KK, ?uszczki JJ, Czuczwar SJ
SIB 1893, a selective mGluR5 receptor antagonist, potentiates the anticonvulsant activity of oxcarbazepine against amygdala-kindled convulsions in rats.
Pol J Pharmacol. 2004 Jul-Aug;56(4):459-64.
SIB 1893 (a non-competitive antagonist of group I metabotropic glutamate receptors), given at 40 mg/kg (but not at 20-30 mg/kg), shortened the afterdischarge duration in amygdala-kindled rats, being ineffective on other seizure parameters - seizure severity, seizure duration, and afterdischarge threshold. Oxcarbazepine (at 7.5 mg/kg, but not at 5 mg/kg), a newer antiepileptic drug, reduced seizure severity, seizure and afterdischarge durations. When combined at ineffective doses in amygdala kindling, SIB 1893 at 20 or 30 mg/kg and oxcarbazepine at 5 mg/kg, significantly reduced seizure and afterdischarge durations. The results indicate that combinations of oxcarbazepine with antagonists of group I metabotropic glutamate receptors may offer a novel therapeutic approach in cases of drug-resistant epilepsy. [Abstract/Link to Full Text]

Folwarczna J, Janiec W, Gawor M, Pytlik M, Kaczmarczyk-Sedlak I, Nowi?ska B
Effects of enoxaparin on histomorphometric parameters of bones in rats.
Pol J Pharmacol. 2004 Jul-Aug;56(4):451-7.
Enoxaparin sodium is a low-molecular-weight heparin. It is not clear whether the risk of development of osteoporosis after administration of low-molecular-weight heparins is lower than after administration of standard heparin. The aim of the present study was to investigate the effects of enoxaparin on histomorphometric parameters of bones in female Wistar rats (13-15 weeks old at the beginning of the experiment). Enoxaparin was administered at doses of 1000 anti-Xa IU/kg sc daily or 2000 anti-Xa IU/kg sc daily for 4 weeks. Bone mass, mineral and calcium content (in the tibia, femur and L-4 vertebra), length and diameter in the tibia and femur, and histomorphometric parameters of the tibia (periosteal and endosteal transverse growth, width of periosteal and endosteal osteoid, area of the transverse cross-section of the cortical bone in the diaphysis and area of the transverse cross-section of the marrow cavity) and the femur (width of epiphyseal and metaphyseal trabeculae, width of epiphyseal cartilage) were examined. Enoxaparin, administered at doses of 1000 anti-Xa IU/kg sc daily or 2000 anti-Xa IU/kg sc daily for 28 days, induced osteopenic changes in the rat skeletal system. The changes observed in bone histomorphometric parameters indicate that enoxaparin caused the inhibition of bone formation and intensification of bone resorption. [Abstract/Link to Full Text]

William BM, Abdel-tawab AM, Hassan EA, Mohamed OF
Acetylator phenotyping in patients with malignant lymphomas, using caffeine as the metabolic probe.
Pol J Pharmacol. 2004 Jul-Aug;56(4):445-9.
OBJECTIVE: The study was designed to answer the question whether a particular (slow/fast) acetylator phenotype is a risk for malignant lymphoma patients. Differences in acetylator phenotypes were previously described in urinary bladder and colorectal carcinoma patients compared to controls. Addressing this issue may help understanding the possible role of xenobiotics' metabolism in the pathogenesis of malignant lymphomas. Caffeine is currently preferred as a metabolic probe due to its noninvasiveness. DESIGN: Case-control study. SETTING: Hematology/oncology inpatient unit and outpatient clinic, Ain Shams University Hospitals, Cairo, Egypt: a tertiary care academic medical institution. PARTICIPANTS: Urine samples were collected 4 h after the oral administration of a caffeine-containing beverage to 83 patients with malignant lymphomas and 92 control subjects. Diagnosis of lymphoma was ascertained histopathologically. Controls were matched in terms of age, sex and residence (urban/rural). MEASUREMENTS: To define the acetylation phenotype, the nanomolar ratio of the urinary concentrations of the caffeine metabolites: acetylamino-6-amino-3-methyluracil and 1-methylxanthine (AAMU/1X) was calculated for every study subject. RESULTS: An excess of fast acetylators was shown in the malignant lymphoma group compared to the control. The odds ratio for malignant lymphoma associated with fast acetylation was 1.304 (95% CI, 0.709-2.398; p = 0.439). Paradoxically, an excess of the slow acetylator phenotype was observed in patients with low-grade non-Hodgkin lymphoma (n = 25) compared to other subjects of the lymphoma group. The odds ratio for low-grade lymphoma associated with slow acetylation was 4.00 (95% CI, 1.316-12.155; p = 0.014). CONCLUSIONS: Owing to the small sample size, the association between the acetylator phenotype and malignant lymphomas could not be excluded. However, the study suggests that, compared to other types of lymphoma, low-grade non-Hodgkin lymphoma would be associated with slow acetylator phenotype. [Abstract/Link to Full Text]

Ozkan A, Fi?kin K
Epirubicin HCl toxicity in human-liver derived hepatoma G2 cells.
Pol J Pharmacol. 2004 Jul-Aug;56(4):435-44.
Epirubicin HCl is a new anthracycline analog and derivative of doxorubicin. Doxorubicin is a potent anticancer agent, the use of which is limited by its cumulative dose-dependent cardiotoxicity. Epirubicin HCl has more favorable therapeutic index than doxorubicin and possesses less hematologic and cardiac toxicity at comparable doses. Hepatoma G2 cells are a valuable model to study hepatocellular carcinoma and the liver, where drugs are metabolized. The goal of our study was to evaluate the cytotoxic effect of epirubicin HCl on viability of Hep G2 cells measured using the MTT cytotoxicity test. Epirubicin HCl produced a concentration- and time-dependent cytotoxicity to Hep G2 cells. The mechanism of cytotoxicity of epirubicin HCl (IC(50) value of 1.6 mug/ml within 24 h) appeared to involve a production of free radical species since activities of free radical scavenging enzymes (SOD, catalase, Se-dependent GPx) were increased. Addition of SOD prevented cytotoxicity of epirubicin HCl, and also counteracted the apoptosis. DNA fragmentation was determined to evaluate apoptosis. Western blot analysis indicated a decrease in GST-pi expression and increased activity of NADPH-dependent cytochrome P450 reductase which is a major enzyme in the conversion of epirubicin HCl to a free radical. It is proposed that production of reactive oxygen species increased by the treatment with epirubicin HCl can cause lipid peroxidation, which subsequently promotes apoptosis and reduces cell viability. Superoxide dismutase, catalase and glutathione peroxidase must be considered as a part of the intracellular antioxidant defense mechanism of Hep G2 cells against single electron reducing quinone-containing anticancer antibiotics. [Abstract/Link to Full Text]

Li Z, Li D, Huang J, Zhang W, Ding Y, Wang S
Preparation of cardiovascular disease-related genes microarray and its application in exploring ligustrazine-induced changes in endothelial gene expression.
Pol J Pharmacol. 2004 Jul-Aug;56(4):427-33.
To study the molecular mechanism of action of ligustrazine, a low-density oligonucleotide microarray for cardiovascular disease-related genes, was constructed, and the preparation and hybridization protocols were optimized. Under the optimized conditions, the molecular mechanism of action of ligustrazine was analyzed with human umbilical vein endothelial cells. After 4 h of treatment with 100 microg/ml of ligustrazine, calcium-ATPase gene, sodium channel gene, P450c11 gene in human umbilical vein endothelial cells were up-regulated while apolipoprotein C-III gene was down-regulated. The results were shown to be reproducible. RT-PCR confirmed the results from microarray. These results suggest that ligustrazine may act on the function of human umbilical vein endothelial cells via modulating the expressions of cardiovascular disease-related genes. This also demonstrates the use of oligonucleotide microarray technology as an approach to studying targets of active components of Traditional Chinese Medicine. [Abstract/Link to Full Text]

Dejda A, Gendek-Kubiak H, Nowak JZ
Effect of different forms of vasoactive intestinal peptide on cAMP formation in the mammalian cerebral cortex.
Pol J Pharmacol. 2004 Jul-Aug;56(4):421-5.
Chicken, guinea pig and mammalian (human/porcine/rat) vasoactive intestinal peptides (VIP; 0.001-3 microM) were compared with respect to their ability to stimulate adenosine 3', 5'-cyclic monophosphate (cAMP) formation in the cerebral cortical slices of rat and guinea pig. Of the tested peptides, i.e. chicken VIP (cVIP), guinea pig VIP (gpVIP) and human/rat/porcine (mammalian) VIP (mVIP), the strongest stimulator of cAMP synthesis was cVIP, and the weakest one - the gpVIP. Pituitary adenylate cyclase-activating polypeptide (PACAP) used as a reference drug at 0.1 microM concentration strongly stimulated cAMP formation in the cerebrum of both species, being, however, significantly more potent in the guinea pig model. The obtained data demonstrate significant differences in biological activity between cVIP and two distinct mammalian VIPs in the cerebral cortex of rat and guinea pig. [Abstract/Link to Full Text]

Recent Articles in Acta Pharmacologica Sinica

Xie SQ, Wu YL, Cheng PF, Wang MW, Liu GC, Ma YF, Zhao J, Wang CJ
A novel homospermidine conjugate inhibits growth and induces apoptosis in human hepatoma cells.
Acta Pharmacol Sin. 2007 Nov;28(11):1827-34.
AIM: To elucidate the mechanism responsible for the antiproliferative effects of a novel homospermidine conjugate, anthracenylmethyl homospermidine (ANTMHspd), in the human hepatoma BEL-7402 cell line. METHODS: The viability of the cells was assessed by MTT assay and the trypan blue dye exclusion method. Morphological changes were observed by fluorescence microscopy with Hoechst 33258 staining. Cell cycle distribution, apoptosis, and mitochondrial membrane potential were measured by flow cytometry. Protein expression was detected by Western blot analysis. RESULTS: ANTMHspd strongly decreased BEL-7402 cell proliferation in a dose- and time-dependent manner. Hoechst 33258 staining and the flow cytometry assay showed that ANTMHspd induced cell apoptosis and cell cycle perturbation. Furthermore, ANTMHspd could induce mitochondrial membrane potential loss and cytochrome c release and enhance cleaved caspase-3, cleaved caspase-9, and Bax protein expression without caspase-8 activation. ANTMHspd could also decrease the expression of Bcl-2 and cytochrome c in mitochondria. In addition, the specific inhibitors of caspase-9 and caspase-3 almost abolished the ANTMHspd-induced caspase-9 and caspase-3 activation, respectively. CONCLUSION: ANTMHspd could induce BEL-7402 cell apoptosis via the mitochondrial/caspase-dependent pathway and the Bcl-2 family was involved in the control of apoptosis. [Abstract/Link to Full Text]

Hu HZ, Yang YB, Xu XD, Shen HW, Shu YM, Ren Z, Li XM, Shen HM, Zeng HT
Oridonin induces apoptosis via PI3K/Akt pathway in cervical carcinoma HeLa cell line.
Acta Pharmacol Sin. 2007 Nov;28(11):1819-26.
AIM: To investigate the apoptosis-inducing effect of oridonin, a diterpenoid isolated from Rabdosia rubescens, in the human cervical carcinoma HeLa cell line. METHODS: A morphological analysis, nuclear condensation, and fragmentation of chromatin were monitored using Hoechst 33342 staining. Cell viability was assessed using the 3-(4, 5-dimethylthiazol-(2)-yl)-2, 5-diphenyl tetrazolium bromide (MTT) assay. Cell apoptosis and the apoptosis-related activation in the HeLa cell line were evaluated by flow cytometry and Western blotting. RESULTS: Oridonin suppressed the proliferation of the HeLa cell line in a dose- and time-dependent fashion. Oridonin treatment downregulated the activation of protein kinase B (Akt), the expression of forkhead box class O (FOXO) transcription factor, and glycogen synthase kinase 3 (GSK3). Oridonin also induced the release of cytochrome c accompanied by the activation of caspase-3 and poly-adenosine diphosphate- ribose polymerase cleavage. In addition, Z-D(OMe)-E(OMe)-V-D(OMe)- FMK (z-DEVD-fmk), an inhibitor of caspases, prevented caspase-3 activation and abrogated oridonin-induced cell death. Finally, oridonin treatment of the HeLa cell line downregulated the expression of the inhibitor of the apoptosis protein. CONCLUSION: Our results showed that oridonin-induced apoptosis involved several molecular pathways. Oridonin may suppress constitutively activated targets of phosphatidylinositol 3-kinase (Akt, FOXO, and GSK3) in the HeLa cell line, inhibiting the proliferation and induction of caspase-dependent apoptosis. [Abstract/Link to Full Text]

Liu BC, Xia HL, Wu JN, Zhang XL, Liu DG, Gong YX
Influence of irbesartan on expression of ILK and its relationship with epithelial-mesenchymal transition in mice with unilateral ureteral obstruction.
Acta Pharmacol Sin. 2007 Nov;28(11):1810-8.
AIM: Irbesartan, a new antagonist of the type 1 angiotensin II receptor, has been proven to be renal protective in both diabetic and non-diabetic nephropathy, but its exact mechanism is still uncertain. Here we investigated the influence of irbesartan on the expression of the integrin-linked kinase (ILK) and its relationship with epithelial-mesenchymal transition (EMT) in mice with unilateral ureteral obstruction (UUO). METHODS: The mice were randomly divided into 3 groups: sham operation (C, n=20), UUO (n=40), and UUO with irbesartan treatment (UUO+irbesartan, n=40). Irbesartan was given at a dose of 50 mg/kg body weight per day by gavage. The experimental animals in the control group received the same volume of vehicle (0.9% saline solution). The animals were sacrificed at d 1, 3, 7, and 14, respectively, after the surgery. RESULTS: The expression of the ILK at mRNA and protein levels were significantly increased in the UUO group 1 d after the surgery, which was significantly decreased by treatment with irbesartan (P<0.01, respectively). The expression of alpha-smooth muscle actin (alpha-SMA) was significantly increased, while E-cadherin was decreased in mice with UUO at d 3 after the surgery. Treatment with irbesartan significantly abrogated such effects (P<0.01). The immunohistochemistry analysis indicated that the protein expression of the ILK was positively correlated with alpha-SMA, but negatively with E-cadherin. CONCLUSION: These results suggested that irbesartan attenuated renal tubulointerstitial fibrosis in UUO mice, which may be related to the inhibition of ILK expression, subsequently preventing the tubular EMT. [Abstract/Link to Full Text]

Wang H, Xu DX, Lu JW, Zhao L, Zhang C, Wei W
N-acetylcysteine attenuates lipopolysaccharide-induced apoptotic liver damage in D-galactosamine-sensitized mice.
Acta Pharmacol Sin. 2007 Nov;28(11):1803-9.
AIM: To investigate the effects of N-acetylcysteine on D-galactosamine (GalN)/ lipopolysaccharide (LPS)-induced apoptotic liver injury in mice. METHODS: When given together with a low dose of LPS, GalN highly sensitizes animals to produce apoptotic liver injury with severe hepatic congestion, resulting in rapid death. In the GalN/LPS model, TNF-alpha is the major mediator leading to apoptotic liver injury. Reactive oxygen species (ROS) are involved in GalN-induced sensitization to TNF-alpha-evoked hepatocyte apoptosis. N-acetylcysteine (NAC) is an antioxidant and a glutathione (GSH) precursor. In this study, we investigated the effects of NAC on LPS-induced apoptotic liver injury in GalN-sensitized mice. RESULTS: Pretreatment with NAC significantly reduced GalN/LPS-induced elevation of serum alanine aminotransferase levels. In parallel, GalN/LPS-induced hepatic necrosis and congestion were obviously improved by NAC. Furthermore, NAC pretreatment significantly alleviated GalN/LPS-induced hepatic apoptosis, measured by the inhibition of hepatic caspase-3 activity and attenuation of DNA laddering. NAC pretreatment had no effect on LPS-evoked nitric oxide production in GalN-sensitized mice. Increases in serum TNF-alpha concentration, which were observed in GalN/LPS-treated mice, were not significantly reduced by NAC. Although NAC pretreatment significantly alleviated LPS-induced hepatic GSH depletion, DL-buthionine-(SR)-sulfoximine, an inhibitor of GSH synthesis, did not influence the protective effect of NAC on GalN/LPS-induced apoptotic liver injury. CONCLUSION: NAC attenuates GalN/LPS-induced apoptotic liver injury via its strong ROS scavenging and anti-apoptotic effects. [Abstract/Link to Full Text]

Guo HS, Cai ZX, Wu TH, Xu J, Qiu Y, Xu WX
Inhibitory effect of dendroaspis natriuretic peptide on spontaneous contraction in gastric antral circular smooth muscles of guinea pigs.
Acta Pharmacol Sin. 2007 Nov;28(11):1797-802.
AIM: To determine whether the natriuretic peptide receptor (NPR) is present in the stomach of guinea pigs and to investigate the effect of dendroaspis natriuretic peptide (DNP) on the gastric motility of guinea pigs and its mechanism. METHODS: The distribution of the NPR was analyzed by autoradioimmunography. The spontaneous contraction of gastric antral circular muscles of guinea pigs was recorded by a 4-channel physiograph. The whole cell patch-clamp technique was introduced to record calcium-activated potassium currents in the gastric myocytes isolated by collagenase. RESULTS: The NPR existed in the gastric fundus, gastric body, and gastric antrum of guinea pigs, and its density was largest in the gastric antrum. DNP inhibited spontaneous contraction and exhibited a dose-dependent manner. The DNP-induced inhibition was diminished by LY83583 (a guanylate cyclase inhibitor) and was potentiated by zaprinast (a cGMP-sensitive phosphoesterase inhibitor). The inhibitory effect of DNP on spontaneous contraction was also inhibited by tetraethylammonium (a non-selective potassium channel blocker); 10 nmol/L DNP increased the calcium-activated potassium currents in the gastric circular myocytes of guinea pigs. CONCLUSION: The NPR is most common in the gastric antrum of guinea pigs. DNP significantly inhibits gastric motility in the gastric antrum of guinea pigs. The inhibitory effect occurs via a cGMP-dependent pathway, and a calcium-activated potassium channel may be also involved in the relaxation induced by DNP in gastric antral circular smooth muscles. [Abstract/Link to Full Text]

Wang K, Shen HH, Li W, Huang HQ
Human C-C chemokine receptor 3 monoclonal antibody inhibits pulmonary inflammation in allergic mice.
Acta Pharmacol Sin. 2007 Nov;28(11):1791-6.
AIM: To evaluate the effect of C-C chemokine receptor 3 (CCR3) blockade on pulmonary inflammation and mucus production in allergic mice. METHODS: We used the synthetic peptide of the CCR3 NH2-terminal as the immunizing antigen and generated murine monoclonal antibody against the human CCR3. In addition, the generated antibody was administered to mice sensitized and challenged with ovalbumin. The inflammatory cells in bronchoalveolar lavage, cytokine levels, pulmonary histopathology, and mucus secretion were examined. RESULTS: The Western blotting analysis indicated that the generated antibody bound to CCR3 specifically. The allergic mice treated with the antihuman CCR3 antibody exhibited a significant reduction of pulmonary inflammation accompanied with the alteration of cytokine. CONCLUSION: The antibody we generated was specific to CCR3. The inhibition of airway inflammation and mucus overproduction by the antibody suggested that the blockade of CCR3 is an appealing therapeutical target for asthma. The present research may provide an experimental basis for the further study of this agent. [Abstract/Link to Full Text]

Zhang HT, Feng ZL, Wu J, Wang YJ, Guo X, Liang NC, Zhu ZY, Ma JQ
Sodium butyrate-induced death-associated protein kinase expression promote Raji cell morphological change and apoptosis by reducing FAK protein levels.
Acta Pharmacol Sin. 2007 Nov;28(11):1783-90.
AIM: To investigate the role of death-associated protein kinase (DAPK) on the apoptosis of Raji cells induced by sodium butyrate. METHODS: The apoptosis of Raji cells were induced by sodium butyrate for 2, 4, 6, 8, and 10 d. Simultaneity, the Raji cells were inhibited to adhere on culture flask by polyHEME. Cell viability was detected by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide method and the cell apoptosis percentage was estimated by flow cytometry. DAPK and focal adhesion kinase (FAK) expression were measured by Western blotting. Coding sequence on the C-terminal of DAPK, which can suppress the function of DAPK, was transfected into the Raji cells to investigate whether the C-terminal of DAPK could inhibit the apoptosis of Raji cells induced by sodium butyrate. RESULTS: After being treated with sodium butyrate, the Raji cells expressed DAPK and displayed many protrusions to adhere onto the culture flask. The Raji cells were susceptible to apoptosis when they were inhibited adhesion by polyHEME. At that time, the cell viability decreased, the cell apoptosis percentage increased and the protein levels of total FAK were reduced. The Raji cells, which were transfected with the coding region on the C-terminal of DAPK, sustained apoptosis and the FAK protein level when treated with sodium butyrate. CONCLUSION: Sodium butyrate induced DAPK expression. It caused the Raji cells to display many protrusions all around the cells and adhere onto the culture flask. DAPK expression prompted apoptosis by reducing the FAK protein level in sodium butyrate-induced Raji cells. [Abstract/Link to Full Text]

Hou Z, Meng JR, Zhao JR, Hu BQ, Liu J, Yan XJ, Jia M, Luo XX
Inhibition of beta-lactamase-mediated oxacillin resistance in Staphylococcus aureus by a deoxyribozyme.
Acta Pharmacol Sin. 2007 Nov;28(11):1775-82.
AIM: To investigate the oxacillin susceptibility restoration of methicillin-resistant Staphylococcus aureus (MRSA) by targeting the signaling pathway of blaR1- blaZ with a DNAzyme. METHODS: A DNAzyme (named PS-DRz602) targeting blaR1 mRNA was designed and synthesized. After DRz602 was introduced into a MRSA strain WHO-2, the colony-forming units of WHO-2 on the Mueller-Hinton agar containing 6 mg/L oxacillin and the minimum inhibitory concentrations of oxacillin were determined. The inhibitory effects of DRz602 on the expressions of antibiotic- resistant gene blaR1 and its downstream gene blaZ were detected by real time RT-PCR. RESULTS: PS-DRz602 significantly decreased the transcription of blaR1 mRNA and led to the significant reduction of blaZ in a concentration-dependent manner. Consequently, the resistance of S aureus WHO-2 to the beta-lactam antibiotic oxacillin was significantly inhibited. CONCLUSION: Our results indicated that blocking the blaR1-blaZ signaling pathway via DNAzyme might provide a viable strategy for inhibiting the resistance of MRSA to beta-lactam antibiotics and that BlaR1 might be a potential target for pharmacological agents combating MRSA. [Abstract/Link to Full Text]

Shang YC, Wang SH, Xiong F, Zhao CP, Peng FN, Feng SW, Li MS, Li Y, Zhang C
Wnt3a signaling promotes proliferation, myogenic differentiation, and migration of rat bone marrow mesenchymal stem cells.
Acta Pharmacol Sin. 2007 Nov;28(11):1761-74.
AIM: To investigate the effects of the wingless-related MMTV integration site 3A (Wnt3a) signaling on the proliferation, migration, and the myogenic and adipogenic differentiation of rat bone marrow mesenchymal stem cells (rMSC). METHODS: Primary MSC were isolated and cultured from Sprague-Dawley rats and characterized by flow cytometry. Mouse L cells were transfected with Wnt3a cDNA, and conditioned media containing active Wnt3a proteins were prepared. Cell proliferation was evaluated by cell count and 5-bromodeoxyuridine incorporation assay. The migration of rMSC was performed by using a transwell migration and wound healing assay. The myogenic and adipogenic differentiation in rMSC were examined by light microscopy, immunofluorescence, and RT-PCR at different time points after myogenic or adipogenic introduction. RESULTS: Wnt3a signaling induced beta-catenin nuclear translocation and activated the Wnt pathway in rMSC. In the presence of Wnt3a, rMSC proliferated more rapidly than the control cells, keeping their differentiation potential. Moreover, Wnt3a signaling induced 2.62% and 3.76% of rMSC-expressed desmin and myosin heavy chain after being cultured in myogenic medium. The myogenic differentiation genes, including Pax7, MyoD, Myf5, Myf4, and myogenin, were activated after Wnt3a treatment. On the other hand, Wnt3a inhibited the adipogenic differentiation in rMSC through the downregulated expression of CCAAT/enhancer-binding protein alpha (C/EBPalpha) and peroxisome proliferator-activated receptor gamma (PPARgamma). Furthermore, Wnt3a promoted the migration capacity of rMSC. CONCLUSION: The results indicate that Wnt3a signaling can induce myogenic differentiation in rMSC. Wnt3a signaling is also involved in the regulation of the proliferation and migration of rMSC. These results could provide a rational foundation for cell-based tissue repair in humans. [Abstract/Link to Full Text]

Ling G, Liu AJ, Shen FM, Cai GJ, Liu JG, Su DF
Effects of combination therapy with atenolol and amlodipine on blood pressure control and stroke prevention in stroke-prone spontaneously hypertensive rats.
Acta Pharmacol Sin. 2007 Nov;28(11):1755-60.
AIM: To test the effects of atenolol and amlodipine, either alone or in combination, on blood pressure, blood pressure variability (BPV), baroreflex sensitivity (BRS), and the prevalence of stroke in stroke-prone spontaneously hypertensive rats (SHR-SP). METHODS: In the first set of the study, 24 8-month-old, female SHR-SP rats were randomly divided into 3 groups. Blood pressure, heart period, and BRS were determined before and after the intragastric administration of atenolol (10 mg/kg) and amlodipine (1.0 mg/kg), either alone or in combination. In the second set of the study, 40 male and 40 female rats were randomly assigned to 1 of the following 4 groups: control, atenolol (10, amlodipine (1.0, and both (10 male and 10 female in each group). The stroke incident and survival time were recorded. RESULTS: Atenolol and amlodipine, either alone or in combination, significantly decreased blood pressure, with the exception of the amlodipine-induced effect on diastolic blood pressure. Meanwhile, only the combination treatment significantly decreased the BPV levels for the same period. The q-values calculated by the probability sum analysis were 1.17 and 2.67 for systolic and diastolic blood pressure, respectively, and were 2.48 and 2.10 for systolic and diastolic BPV, respectively, following administration. Neither drug exhibited any significant effect on BRS. Atenolol and amlodipine, either alone or in combination, significantly increased the lifespan of SHR-SP, with the best effect elicited by the combination therapy. CONCLUSION: A significant synergism exists between atenolol and amlodipine in lowering and stabilizing blood pressure in SHR-SP. Combination therapy may be an optimal strategy for the prevention of stroke in hypertension. [Abstract/Link to Full Text]

Feng Y, Tang XY, Dai DZ, Dai Y
Reversal of isoproterenol-induced downregulation of phospholamban and FKBP12.6 by CPU0213-mediated antagonism of endothelin receptors.
Acta Pharmacol Sin. 2007 Nov;28(11):1746-54.
AIM: The downregulation of phospholamban (PLB) and FKBP12.6 as a result of beta- receptor activation is involved in the pathway(s) of congestive heart failure. We hypothesized that the endothelin (ET)-1 system may link to downregulated PLB and FKBP12.6. METHODS: Rats were subjected to ischemia/reperfusion (I/R) to cause heart failure (HF). 1 mg/kg isoproterenol (ISO) was injected subcutaneously (sc) for 10 d to worsen HF. 30 mg/kg CPU0213 (sc), a dual ET receptor (ETAR/ETBR) antagonist was given from d 6 to d 10. On d 11, cardiac function was assessed together with the determination of mRNA levels of ryanodine receptor 2, calstabin-2 (FKBP12.6), PLB, and sarcoplasmic reticulum Ca2+-ATPase. Isolated adult rat ventricular myocytes were incubated with ISO at 1X10(-6) mol/L to set up an in vitro model of HF. Propranolol (PRO), CPU0213, and darusentan (DAR, an ETAR antagonist) were incubated with cardiomyocytes at 1X10(-5) mol/L or 1X10(-6) mol/L in the presence of ISO (1X10(-6) mol/L). Immunocytochemistry and Western blotting were applied for measuring the protein levels of PLB and FKBP12.6. RESULTS: The worsened hemodynamics produced by I/R were exacerbated by ISO pretreatment. The significant downregulation of the gene expression of PLB and FKBP12.6 and worsened cardiac function by ISO were reversed by CPU0213. In vitro ISO 1X10(-6) mol/L produced a sharp decline of PLB and FKBP12.6 proteins relative to the control. The downregulation of the protein expression was significantly reversed by the ET receptor antagonist CPU0213 or DAR, comparable to that achieved by PRO. CONCLUSION: This study demonstrates a role of ET in mediating the downregulation of the cardiac Ca2+-handling protein by ISO. [Abstract/Link to Full Text]

Li XG, Yan JT, Xu XZ, Wang JN, Cheng LM, Wang T, Zuo P, Wang DW
Recombinant adeno-associated virus-mediated delivery of antisense angiotensin II receptor 1 gene attenuates hypertension development.
Acta Pharmacol Sin. 2007 Nov;28(11):1737-45.
AIM: The renin-angiotensin system plays a crucial role in the development and establishment of hypertension, and the pharmacological blockade of the system results in a reduction in blood pressure. In the present study, we investigated whether the effects of a novel, double-stranded, recombinant adeno-associated virus vector (rAAV)-mediated antisense angiotensin II receptor 1 (AT1R) gene efficiently prevents the development of hypertension induced by a high-salt diet in adult, male Sprague-Dawley (SD) rats. METHODS: A rAAV was prepared with a cassette containing a cytomegalovirus promoter and partial cDNA (660 base pairs) for the AT1R inserted in the antisense direction (rAAV-AT1-AS). A single tail vein injection of the rAAV-AT1-AS or rAAV-GFP (green fluorescent protein, a reporter gene) was performed in adult, male SD rats. Two weeks after injection, the animals were fed a diet containing 8% NaCl, and the systolic blood pressure was measured weekly using the tail-cuff method for 12 weeks. RESULTS: The high-salt diet induced a significant rise in systolic blood pressure in the rAAV-GFP-treated animals; however, the rAAV-AT1-AS treatment attenuated the rise in blood pressure (142.7+/-4.5 mmHg vs 117+/-3.8 mmHg, P<0.01), and the hypotensive effect was maintained until the experiments ended at 12 weeks. In the rAAV-GFP-treated animals AT1 was overexpressed in various tissues, especially in the aorta and kidney at mRNA levels; in contrast, rAAV-AT1-AS treatment markedly attenuated AT1 expression. Furthermore, rAAV-AT1-AS treatment prevented target organ damages from hypertension, including cardiac dysfunction and renal injury compared to the rAAV-GFP group. CONCLUSION: These results suggest that rAAVmediated anti-AT1 delivery attenuates the development of hypertension and protects against renal injury and cardiac remodeling. [Abstract/Link to Full Text]

Weng XC, Zheng JQ, Li J, Xiao WB
Underlying mechanism of ASIC1a involved in acidosis-induced cytotoxicity in rat C6 glioma cells.
Acta Pharmacol Sin. 2007 Nov;28(11):1731-6.
AIM: To investigate the underlying mechanism of acid-sensing ion channel (ASIC) 1a involved in the acidosis-induced cytotoxicity of rat C6 glioma cells. METHODS: The stable ASIC1a-silenced C6 cells built with the RNA interference technology were confirmed by RT-PCR and Western blot analysis. Intracellular calcium ([Ca2+]i) in both the wild-type rat C6 glioma cells and the ASIC1a-silenced C6 cells were analyzed before and after acid application/exposure with the calcium imaging experiment. RESULTS: The rapid extracellular pH drop induced the increase of [Ca2+]i in the wild-type C6 cells, but not in the ASIC1a-silenced C6 cells. During the prolonged acid exposure, [Ca2+]i was lower in the ASIC1a-silenced C6 cells than that in the control cells. CONCLUSION: The resultant toxicity of [Ca2+]i might contribute to the acidosis-induced cytotoxicity. [Abstract/Link to Full Text]

Gong QH, Wang Q, Shi JS, Huang XN, Liu Q, Ma H
Inhibition of caspases and intracellular free Ca2+ concentrations are involved in resveratrol protection against apoptosis in rat primary neuron cultures.
Acta Pharmacol Sin. 2007 Nov;28(11):1724-30.
AIM: To investigate the influence of resveratrol (Res), a nutritional antioxidant, on the inhibition of apoptosis in rat primary neuron cultures. METHODS: The cultured cortical neurons of neonatal Sprague-Dawley rats were pretreated with Res (0.1, 1.0, and 10.0 micromol/L) and oxygen-glucose deprivation/reperfusion (OGD/RP) with oxygen and glucose were initiated at d 10 in vitro. Neuronal apoptosis was determined by flow cytometry, and morphological changes of neurons were observed by an electron microscope. For the mechanism studies, the intracellular free calcium concentration ([Ca2+]i) and the transcription of caspases-3 and -12 in neurons were detected by Fura 2/AM loading and real-time RT-PCR, respectively. RESULTS: OGD/RP insult could induce an increase in the apoptotic rate of neurons (from 11.1% to 49.0%), and elicit an obvious morphological change in neurons; pretreatments with Res (0.1, 1.0, and 10.0 micromol/L, respectively) significantly reduced the elevated rate of apoptosis to 41.7%, 40.8%, and 37.4%, respectively, and ameliorated the neuronal morphological injury. Similarly, the OGD/RP insult obviously elicited the elevated levels of the [Ca2+]i and the expressions of caspases-3 and -12 mRNA in neurons. Res pretreatments markedly depressed the neuronal abnormal elevation of [Ca2+]i and the overexpression of caspases-3 and -12 mRNA in a concentration-dependent manner. CONCLUSION: Res can attenuate the rat cortical neuronal apoptosis induced by OGD/RP. The mechanisms are, at least partly, due to the inhibition of the calcium overload and the overexpression of caspases-3 and -12 mRNA. [Abstract/Link to Full Text]

Fang J, Liu R, Tian Q, Hong XP, Wang SH, Cao FY, Pan XP, Wang JZ
Dehydroevodiamine attenuates calyculin A-induced tau hyperphosphorylation in rat brain slices.
Acta Pharmacol Sin. 2007 Nov;28(11):1717-23.
AIM: This study was to investigate the effect of dehydroevodiamine (DHED) on Alzheimer's disease (AD)-like tau hyperphosphorylation induced by calyculin A (CA), an inhibitor of protein phosphatase (PP)-2A and PP-1, and the involvement of PP-2A in metabolically competent rat brain slices. METHODS: Rat brain slices were pre-incubated at 33 degree centigrade in the presence (10, 100, and 200 micromol/L, respectively) or absence of DHED for 1 h. Then, CA 0.1 micromol/L was added and the slices were treated for another 2 h. Western blotting and/or immunohistochemistry were used to measure the phosphorylation level of tau and PP-2A. RESULTS: CA treatment could remarkably increase the immunoreactivity of pS262 and decrease the staining of Tau-1, representing tau hyperphosphorylation at Ser262 (pS262) and Ser198/ 199/202 (Tau-1, as the antibody reacts with unphosphorylated tau, therefore, decreased staining represents increased phosphorylation). Pre-incubation of the brain slices with DHED could efficiently attenuate the CA-induced tau hyperphosphorylation at the above AD-related sites. Additionally, DHED also decreased the basal phosphorylation level of tau at Ser396, although CA failed to induce tau hyperphosphorylation at this site. Furthermore, CA treatment induced an increased level of Tyr307-phosphorylated PP-2A, which represents inactivation of the phosphatase, whereas DHED arrested the elevation of the inhibitory modification of PP-2A. CONCLUSION: DHED can attenuate CA-induced tau hyperphosphorylation at multiple AD-related sites in metabolically active rat brain slices. The underlying mechanism may involve a decreased inhibitory phosphorylation of PP-2A at Tyr307. [Abstract/Link to Full Text]

Chen CQ, Xin H, Zhu YZ
Hydrogen sulfide: third gaseous transmitter, but with great pharmacological potential.
Acta Pharmacol Sin. 2007 Nov;28(11):1709-16.
Hydrogen sulfide (H2S), which is well known traditionally as a toxic gas, has been proven to be produced endogenously by 3 enzymes in mammalian tissues and plays important roles in physiological and pathophysiological conditions. In the central nervous system, H2S functions as not only a neuromodulator, but also a neuroprotectant against oxidative stress. In the cardiovascular system, H2S relaxes vascular smooth muscles by the activation of KATP channels and inhibits smooth muscle cell proliferation via the mitogen-activated protein kinase signaling pathway. These effects are important for maintaining blood pressure and preventing vessel structural remodeling, and identifies H2S as an important factor in the development of some vascular diseases, such as hypertension. H2S also shows cardioprotective effects in ischemic myocardium and septic and endotoxin shock. Recent studies have demonstrated a new mechanism to explain the motor effect of H2S on the rat detrusor muscle, which is through the activation of the capsaicin-sensitive primary neuron. This review focuses on the recent research achievements on H2S and discloses the great potential of H2S as the third gaseous transmitter in cardiac protection. [Abstract/Link to Full Text]

Wang J, Xie X
Development of a quantitative, cell-based, high-content screening assay for epidermal growth factor receptor modulators.
Acta Pharmacol Sin. 2007 Oct;28(10):1698-704.
AIM: To develop a robust, cell-based, high-content screening (HCS) assay based on receptor internalization for the identification of novel modulators of the epidermal growth factor receptor (EGFR). METHODS: Agonist-induced receptor internalization is part of the signaling cascade of EGFR. Fluorescent-tagged epidermal growth factor (EGF) was used to visualize the internalized receptor-ligand complex. The fluorescent intracellular spots were detected and measured with an ArrayScan HCS reader. Compounds that can competitively bind to EGFR or interfere with EGFR internalization process would result in a reduced number and intensity of intracellular fluorescent spots. This assay was validated, optimized, and applied to a large-scale screening of a library containing 48,000 synthetic compounds. RESULTS: The competition between fluorescent EGF and unlabeled EGF reveals the IC(50) of unlabeled EGF is approximately 0.2 nmol/L, which is comparable with other published reports. Thirteen compounds with a relatively high degree of interference with EGFR internalization were identified. One of the compounds was proven to be agonist of the EGFR since it induced phosphorylation of the receptor and extracellular signal-regulated protein kinase (ERK). CONCLUSION: This automated, objective, and easy-to-use assay provided abundant information, quantitative results, and demonstrated the potential use of HCS methods in searching membrane receptor modulators. [Abstract/Link to Full Text]

Xu LY, He YJ, Zhang W, Deng S, Li Q, Zhang WX, Liu ZQ, Wang D, Huang YF, Zhou HH, Sun ZQ
Organic anion transporting polypeptide-1B1 haplotypes in Chinese patients.
Acta Pharmacol Sin. 2007 Oct;28(10):1693-7.
AIM: To detect 388G>A and 521T>C variant alleles in the organic anion transporting polypeptide-1B1 (OATP1B1, encoding gene SLCO1B1) gene. METHODS: One hundred and eleven healthy volunteers were screened for OATP1B1 alleles in our study. PCR-restriction fragment length polymorphism was used to identify the 388G>A polymorphism and a 1-step tetra-primer method was developed for the determination of 521T>C mutation. RESULTS: The frequencies of the 388G>A and 521T>C variant alleles in the Chinese population were 73.4% and 14.0%, respectively. The frequencies of the SLCO1B1*1b and *15 haplotypes were 59.9% and 14.0%, respectively. CONCLUSION: The SLCO1B1*1b and SLCO1B1*15 variants are relatively common in the Chinese population. Their frequencies are similar to that in the Japanese, but significantly different from that in Caucasians and blacks. [Abstract/Link to Full Text]

Fan L, Wang G, Wang LS, Chen Y, Zhang W, Huang YF, Huang RX, Hu DL, Wang D, Zhou HH
Herbal medicine yin zhi huang induces CYP3A4-mediated sulfoxidation and CYP2C19-dependent hydroxylation of omeprazole.
Acta Pharmacol Sin. 2007 Oct;28(10):1685-92.
AIM: To explore the potential interactions between yin zhi huang (YZH) and omeprazole, a substrate of CYP3A4 and CYP2C19. METHODS: Eighteen healthy volunteers, including 6 CYP2C19*1/*1, 6 CYP2C19*1/*2 or *3 and 6 CYP2C19*2/*2 were enrolled in a 2-phase, randomized, crossover clinical trial. In each phase, the volunteers received either placebo or 10 mL YZH oral liquid, 3 times daily for 14 d. Then all the patients took a 20 mg omeprazole capsule orally. Blood samples were collected up to 12 h after omeprazole administration. Plasma concentrations of omeprazole and its metabolites were quantified by HPLC with UV detection. RESULTS: After 14 d of treatment of YZH, plasma omeprazole significantly decreased and those of omeprazole sulfone and 5-hydroxyomeprazole significantly increased. The ratios of the area under the plasma concentration-time curves from time 0 to infinity (AUC(0-infinity) of omeprazole to 5-hydroxyomprazole and those of omeprazole to omeprazole sulfone decreased by 64.80%+/-12.51% (P=0.001) and 63.31%+/-18.45% (P=0.004) in CYP2C19*1/*1, 57.98%+/-14.80% (P=0.002) and 54.87%+/-18.42% (P=0.003) in CYP2C19*1/*2 or *3, and 37.74%+/-16.07% (P=0.004) and 45.16%+/-15.54% (P=0.003) in CYP2C19*2/*2, respectively. The decrease of the AUC(0-infinity) ratio of omeprazole to 5-hydroxyomprazole in CYP2C19*1/*1 and CYP2C19*1/*2 or *3 was greater than those in CYP2C19*2/*2 (P=0.047 and P=0.009). CONCLUSION: YZH induces both CYP3A4-catalyzed sulfoxidation and CYP2C19-dependent hydroxylation of omeprazole leading to decreases in plasma omeprazole concentrations. [Abstract/Link to Full Text]

Jiang DC, Wang L, Wang YQ, Li L, Lu W, Bai XR
Population pharmacokinetics of valproate in Chinese children with epilepsy.
Acta Pharmacol Sin. 2007 Oct;28(10):1677-84.
AIM: The aim of the present study is to establish a population pharmacokinetic (PPK) model of valproate (VPA) in Chinese epileptic children to promote the reasonable use of anti-epileptic drugs. METHODS: Sparse data of VPA serum concentrations from 417 epileptic children were collected. These patients were divided into 2 groups: the PPK model group (n=317) and the PPK valid group (n=100). The PPK parameter values of VPA were calculated by NONMEM software using the data of the PPK model group. A basic model and a final model were set up. To validate the 2 models, the concentrations of PPK valid group were predicted by each model, respectively. The mean prediction error (MPE), mean squared prediction error (MSPE), root mean squared prediction error (RMSPE), weight residues (WRES), and the 95% confidence intervals (95% CI) were also calculated. Then, the values between the 2 models were compared. RESULTS: The PPK of VPA was determined by a 1-compartment model with a first-order absorption process. The basic model was: Ka=3.09 (h(-1)), V/F=20.4 (L), CL/F=0.296 (L/h). The final model was: Ka=0.251+2.24 x (1-HS) (h(-1)), V/F=2.88+0.157 x WT (L), CL/F=0.106(0.98 x CO)+ 0.0157 x AGE (L/h). For the basic model, the MPE, MSPE, RMSPE, WRES, and the 95% CI were -23.53 (-30.36, -16.70), 3728.96 (2872.72, 4585.20), 39.62 (34.34, 44.90), and -0.06 (-0.14, 0.02), respectively. For the final model, the MPE, MSPE, RMSPE, WRES, and the 95% CI were -1.16 (-4.85, 2.53), 1002.83 (1050.64, 1143.61), 23.04 (21.12, 24.96), and 0.08 (-0.04, 0.20), respectively. The final model was more optimal than the basic model. CONCLUSION: The PPK model of VPA in Chinese epileptic children was successfully established. It will be valuable to facilitate individualized dosage regimens. [Abstract/Link to Full Text]

Cui LY, Liu SL, Ding Y, Huang DS, Ma RF, Huang WG, Hu BS, Pan QH
IL-1beta sensitizes rat intervertebral disc cells to Fas ligand mediated apoptosis in vitro.
Acta Pharmacol Sin. 2007 Oct;28(10):1671-6.
AIM: To determine the apoptotic effect of recombinant rat Fas Ligand on rat intervertebral disc cells pre-treated with IL-1beta in vitro, and the expression of Fas in cultured rat intervertebral disc cells. METHODS: Cells were isolated from the inner annulus fibrosus and transition zones of lumbar discs from Sprague-Dawley rats. The cells were grown in monolayer and divided in 5 treatment groups. IL-1beta (10 ng/mL), FasL (5, 20 ng/mL) with/without IL-1beta (10 ng/mL) pre-treatment was respectively added in Dulbeccoos modified Eagleos medium and Hamos F-12 medium with 1% fetal bovine serum. After 32 h, the cells were stained with annexin V-FITC and propidium iodide to evaluate apoptosis using flow cytometry and to analysis transcription of Fas using RT-PCR. RESULTS: Compared with control group, FasL (20 ng/mL), IL-1beta (10 ng/mL)+FasL (5 ng/mL), and IL-1beta (10 ng/mL)+FasL (20 ng/mL) induced significant apoptosis of the disc cells (P<0.01). Apoptosis was also induced by FasL 5 ng/mL (P<0.05); whereas, apoptosis was not induced by IL-1beta (10 ng/mL) (P>0.05). IL-1beta (10 ng/mL) enhanced the apoptosis-inducing effects of FasL (5 ng/mL) and FasL (20 ng/mL) in disc cells. Fas gene transcription in all groups and Fas expression in the 5 treatment groups were approximately 1.2-2.1-fold greater than control group (respectively, P<0.05). Additionally, Fas expression in FasL with IL-1beta pre-treatment groups were significantly up-regulated than in FasL groups (P<0.01). CONCLUSION: The results of this study showed disc cells pre-treated with IL-1beta increased apoptotic rate in response to FasL in vitro and provided insights to understand Fas/FasL system-mediated apoptosis in disc cells which would be enhanced due to inflammation factor in degenerative disc. [Abstract/Link to Full Text]

Chen HY, Chu X, Yan CL, Chen XH, Sun M, Wang YJ, Wang CB, Yu WG
Polypeptide from Chlamys farreri attenuates murine thymocytes damage induced by ultraviolet B.
Acta Pharmacol Sin. 2007 Oct;28(10):1665-70.
AIM: Polypeptide from Chlamys farreri (PCF, molecular mass is 879) is a new marine polypeptide compound isolated from Chlamys farreri. This study investigates the possible protective roles and the mechanism of PCF against ultraviolet B (UVB)-induced apoptosis in murine thymocytes. METHODS: The rate of apoptosis and caspase-3 activation was measured by flow cytometry. The expression of stress-response genes c-fos and c-jun was observed by RT-PCR. Western blot analysis was performed to determine the release of cytochrome c. RESULTS: It was found that UVB induced murine thymocyte death. The cells treated with UVB showed an increase in cytochrome c release, caspase-3 activity, as well as in the expression of c-fos and c-jun. In addition, all were involved in UVB-induced cell apoptosis. CONCLUSION: Our present observations pointed to the ability of PCF to avert UVB-induced apoptosis in thymocytes by modulating c-fos and c-jun expression, cytochrome c release, and the consequent activation of caspase-3, which were essential components of the UV-induced cell apoptotic pathway. The results suggested that PCF is a promising protective substance against UV radiation. [Abstract/Link to Full Text]

Lei P, He Y, Ye Q, Zhu HF, Yuan XM, Liu J, Xing W, Wu S, Dai W, Shen X, Wang GB, Shen GX
Antigen-binding characteristics of AbCD71 and its inhibitory effect on PHA-induced lymphoproliferation.
Acta Pharmacol Sin. 2007 Oct;28(10):1659-64.
AIM: To investigate the anti-lymphoproliferative effect of the prepared recombinant chimeric human/murine anti-cluster of differentiation(CD) 71 monoclonal antibody (AbCD71), which is composed of mouse-derived, antigen-binding variable regions and human-derived constant regions. METHODS: After plasmids construction and transfection, the expression of AbCD71 in the transfectoma supernatant was determined by the sandwich ELISA. Indirect immunofluorescence assay was used to measure the antigen-binding characteristic and the percent CD71-expressed peripheral blood mononuclear cells (PBMC). The antibodies were purified from the ascites via diethylaminoethyl(DEAE)-Sephadex A-50 chromatography and then identified by SDS-PAGE. At last, inhibitory effect of AbCD71 on PHA-induced PBMC proliferation was calculated by methyl thiazolyl tetrazolium(MTT) assay. RESULTS: Constant domain of heavy chain (C(H)) and light chain (C(L)) of AbCD71 were in the human Cgamma family and human Ckappa family, respectively. AbCD71 could compete with its original murine mAb to bind with CD71-positive human leukemia cell line CEM cells. AbCD71 could inhibit the peripheral blood mononuclear cell proliferation induced by phytohemagglutinin(PHA) in vitro in a dose-dependent manner, especially at time-points 0 and 12 h after induction. There was no statistical difference when compared with original murine mAb. CONCLUSION: The AbCD71 is a promising immunosuppressant. Our approach to blocking CD71 with the chimeric human/murine mAb provides a novel strategy for prolonging allograft survival. [Abstract/Link to Full Text]

Chen ZY, Cheng AC, Wang MS, Xu DW, Zeng W, Li Z
Antiviral effects of PNA in duck hepatitis B virus infection model.
Acta Pharmacol Sin. 2007 Oct;28(10):1652-8.
AIM: To study the efficacy of antiviral treatment with PNA for the duck model of HBV (DHBV)-infected ducks. PNA is a 2-amine-9-(2,3-dideoxy-2,3-dihydro-beta-D-arabinofuranosyl)-6-methoxy-9H-purine. METHODS: The Sichuan Mallard ducklings in the hepatitis B virus model were treated with PNA, a new antiviral agent. DHBV DNA from the blood serum and liver tissues were measured at 0, 5, and 10 d during the treatment and at 3 d withdrawal by real-time PCR. The duck hepatitis B surface antigen (DHBsAg) in the liver cells was observed by Immunohistochemistry (IHC). Pathological changes in the liver tissues were also observed. Control group I was administered with distilled water and control group II was administered with 3-thiacytidine. Treatment group I was administered with PNA at a dose of 40 mg/kg and treatment group II was administered perorally (po) with PNA at a dose of 80 mg/kg. Treatment group III was administered with PNA at a dose of 20 mg/kg and treatment group IV was intravenously administered with PNA at a dose of 40 mg/kg. Each group contained 15 ducklings. RESULTS: PNA can significantly lower the DHBV replication levels in serum and liver. Compared with control group II, there were no significant differences in inhibiting efficacy in treatment groups I and III (P>0.05) and there were significant differences in inhibiting efficacy in treatment groups II and IV (P<0.05). Interestingly, significant differences were observed at 3 d withdrawal. The DHBV replication levels in each group slightly increased at 3 d withdrawal, but rebounded slightly in the PNA treatment groups than in control group II (P<0.05). The DHBV replication levels in the treatment groups were lower than in control group I. The DHBV replication levels in sera had a positive relationship with that in the liver, but the DHBV replication levels in the liver was lower than that in sera. Pathological changes in the treatment groups were obviously improved and the changes were associated with liver viral DNA levels. CONCLUSION: The results demonstrate that PNA is a strong inhibitor of DHBV replication in the DHBV-infected duck model. [Abstract/Link to Full Text]

Jin CY, Lee JD, Park C, Choi YH, Kim GY
Curcumin attenuates the release of pro-inflammatory cytokines in lipopolysaccharide-stimulated BV2 microglia.
Acta Pharmacol Sin. 2007 Oct;28(10):1645-51.
AIM: Pro-inflammatory mediators, such as prostaglandin E2 (PGE2) and nitric oxide (NO), and pro-inflammatory cytokines such as interleukin (IL)-1beta, IL-6, and TNF-alpha, play pivotal roles in brain injuries. The anti-inflammatory properties are known to be associated with significant reductions in pro-inflammatory mediators in brain injuries. In the present study we investigate whether the effects of curcumin on the production of pro-inflammatory mediators in lipopolysaccharide (LPS)-stimulated BV2 microglia. METHODS: Curcumin were administered and their effects on LPS-induced pro-inflammatory mediators were monitored by Western blotting and RT-PCR. RESULT: Curcumin significantly inhibited the release of NO, PGE2, and pro-inflammatory cytokines in a dose-dependent manner. Curcumin also attenuated the expressions of inducible NO synthase and cyclooxygenase-2 mRNA and protein levels. Moreover, curcumin suppressed NF-kappaB activation via the translocation of p65 into the nucleus. Our data also indicate that curcumin exerts anti-inflammatory properties by suppressing the transcription of proinflammatory cytokine genes through the NF-kappaB signaling pathway. CONCLUSION: Anti-inflammatory properties of curcumin may be useful for treating the inflammatory and deleterious effects of microglial activation in response to LPS stimulation. [Abstract/Link to Full Text]

Du HY, Xia SY, Song HF, Li N, Shang MM, Zou J, Wang QQ, Ou L, Sun X, Ji AG, Tang ZM
Structure-efficacy relationships of immunostimulatory activity of CpG-containing oligodeoxynucleotides on mouse spleen cells.
Acta Pharmacol Sin. 2007 Oct;28(10):1637-44.
AIM: To study the relationship between primary structures of oligodeoxynucleotides (ODN) containing unmethylated deoxycytidyldeoxyguanosine (CpG) dinucleotide motifs and their immunostimulatory activities in mouse spleen cells. METHODS: A series of CpG ODN with different primary structures were synthesized. Their capabilities to stimulate mouse spleen cell proliferation were determined by [3H]thymidine incorporation assay. Cytokine (interleukin [IL]-6, IL-12, and IFN-alpha) secretion spectra induced by CpG ODN were assessed by ELISA. The ability of CpG ODN to activate natural killer cells was evaluated by standard 4 h (51)Cr-release assay. Flow cytometry was utilized to examine the expressions of various lymphocyte surface molecules on diverse immunocytes. An effective CpG ODN for murine, ODN1826, was set as the template of modification and the positive control. RESULTS: The immunostimulatory activities of CpG ODN with different sequences and compositions varied markedly, both in character and in extent. It was useless for improving the immunostimulatory activity of ODN1826 by simply increasing the functional hexameric CpG motif number, modifying the site of CpG motifs, or changing the distance between multi-CpG motifs. However, an addition of a self-complementary palindrome structure at the 3'-end, but not the 5'-end of CpG ODN, aroused marked improvement in its activity. Several designed ODN had superior comprehensive immunostimulatory properties compared to ODN1826. CONCLUSION: The immunostimulatory activity of a CpG ODN was relevant to its primary structure. It was useless for promoting immunostimulatory activity to simply change CpG motif number, space, or distance. The 3'-end palindrome structure of CpG ODN is associated with enhanced immunostimulatory activity. [Abstract/Link to Full Text]

Jiao Y, Ge CM, Meng QH, Cao JP, Tong J, Fan SJ
Adenovirus-mediated expression of Tob1 sensitizes breast cancer cells to ionizing radiation.
Acta Pharmacol Sin. 2007 Oct;28(10):1628-36.
AIM: To investigate the effect of the Tob1 gene, a member of the Transducing Molecule of ErbB2/B-cell Translocation Ggene (TOB/BTG) family, by using the adenovirus-mediated expression of Tob1 on radiosensitivity in a human breast cancer cell line MDA-MB-231. METHODS: Cell survival was determined by clonogenic assay. Apoptosis was evaluated by DNA fragmentation gel electrophoresis and terminal deoxynucleotidyl transferase-mediated nick end labeling assay. Protein expression was analyzed by Western blot assay and DNA repair was measured by a host cell reactivation assay. RESULTS: We demonstrated that pre-irradiation treatment with Ad5-Tob1 significantly increased radiosensitivity, accompanying the increased induction of apoptosis and the repression of DNA damage repair. Furthermore, Ad5-Tob1-mediated radiosensitivity correlates with the upregulation of the pro-apoptotic protein Bax and the downregulation of several DNA double strand break repair proteins, including DNA-dependent protein kinases, Ku70 and Ku80, and X-ray-sensitive complementation group 4. CONCLUSION: Tob1, as a new radiosensitizer, is a new target in the radiotherapy of breast cancer via increasing apoptosis and suppressing DNA repair. [Abstract/Link to Full Text]

Weng XC, Zheng JQ, Jin QE, Ma XY
Inhibition of acid-induced apoptosis by targeting ASIC1a mRNA with short hairpin RNA.
Acta Pharmacol Sin. 2007 Oct;28(10):1621-7.
AIM: To study the role of acid-sensing ion channel (ASIC) 1a in the cell death and apoptosis induced by extracellular acid in C6 glioma cells. METHODS: The stable ASIC1a-silenced C6 cell line, built with RNA interference technology, were confirmed by RT-PCR and Western blot analysis. The cell viability following acid exposure was analyzed with lactate dehydrogenase (LDH) and 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. The apoptotic cells dyed with Annexin-V and propidium iodide were measured with a flow cytometer, while the changes of cell cycle were also assayed. RESULTS: The downregulation of ASIC1a proteins by stable transfection of short hairpin RNA decreased the cell death percentage and increased cell viability following acid exposure with LDH and the MTT assay. The rate of apoptosis was lower in the ASIC1a-silenced cell line than that in the wild-type C6 cell line. The percentage of sub-G0 cells was lower in the ASIC1a-silenced C6 cells than that in the wild-type cells. CONCLUSION: Extracellular acid induced cell death and apoptosis via ASIC1a mechanisms in the C6 glioma cells. [Abstract/Link to Full Text]

Shi M, Zhang L, Gu HT, Jiang FQ, Qian L, Yu M, Chen GJ, Luo Q, Shen BF, Guo N
Efficient growth inhibition of ErbB2-overexpressing tumor cells by anti-ErbB2 ScFv-Fc-IL-2 fusion protein in vitro and in vivo.
Acta Pharmacol Sin. 2007 Oct;28(10):1611-20.
AIM: To investigate the antitumor activities of an anti-ErbB2 scFv-Fc-interleukin 2 (IL-2) fusion protein (HFI) in vitro and in vivo. METHODS: Fusion protein HFI was constructed. The efficacy of HFI in mediating tumor cell lysis was determined by colorimetric lactate dehydrogenase release assays. The antitumor activity of HFI was evaluated in tumor xenograft models. RESULTS: The fusion protein was folded as a homodimer formed by covalently linking Fc portions and it retained ErbB2 specificity and IL-2 biological activity. HFI mediated antibody-dependent cell-mediated cytotoxicity (ADCC) at low effector-to-target ratios in vitro and improved the therapeutic efficacy of IL-2 in experiments in vivo. CONCLUSION: The genetically-engineered anti-ErbB2 scFv-Fc-IL-2 fusion protein exhibited high efficiency both in mediating ADCC in vitro and significant antitumor activity in tumor xenograft models. [Abstract/Link to Full Text]

Zhang C, Tang TT, Ren WP, Zhang XL, Dai KR
Inhibiting wear particles-induced osteolysis with doxycycline.
Acta Pharmacol Sin. 2007 Oct;28(10):1603-10.
AIM: To study the effect of doxycycline (DOX) on osteoclastogenesis, mature osteoclast fate and function, wear particles-induced osteoeolysis, and to provide some foundation for treating aseptic loosening and osteolysis after joint arthroplasty. METHODS: Osteoclasts were generated from mouse bone marrow monocytes with the receptor activator of NF-kappaB ligand and the macrophage colony stimulating factor. DOX at a concentration of 5, 10, 15, and 20 microg/mL was respectively added to the medium. Seven days later, the osteoclasts were determined through tartrate-resistant acid phosphatase (TRAP) staining. Mature osteoclasts were isolated from newborn rabbits and cultured for 3 d in 24-well plates or on bone slices. DOX at a concentration of 5, 10, 15, and 20 microg/mL was respectively added to the medium. After TRAP staining, the osteoclasts were counted, resorption on bone slices was quantified, and the area was calculated after toluidine blue and Mayer-hematoxylin staining. Polymethyl methacrylate (PMMA) or ultra-high molecular weight polyethylene (UHMWPE) particles were implanted on the calvariae of C57BL/J6 mice. DOX, at a dose of 2 and 10 mg x kg(-1) x d(-1), was respectively given intraperitoneally for 7 d. Seven days later, the calvariae were removed and processed for pathological analysis. RESULTS: DOX treatment effectively inhibited in vitro osteoclastogenesis, affected the fate of mature osteoclasts, and inhibited mature osteoclasts, causing bone resorption. In vivo data indicated that DOX strongly inhibited PMMA or UHMWPE-induced osteolysis and osteoclastogenesis. CONCLUSION: DOX can effectively inhibit osteoclastogenesis and affect mature osteoclast fate and suppress wear particles induced by osteolysis and osteoclastogenesis. DOX might be useful in the treatment or prevention of wear particles-induced osteolysis and aseptic loosening for its effect on osteoclast generation and mature osteoclast fate and function. [Abstract/Link to Full Text]

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Population pharmacokinetics and proton pump inhibitory effects of intravenous lansoprazole in healthy Japanese males.
Biol Pharm Bull. 2007 Dec;30(12):2238-43.
A total of 56 healthy Japanese males were enrolled in single- or multiple- dose pharmacokinetic trials of intravenous lansoprazole administration. The population pharmacokinetics of the drug was evaluated using nonlinear mixed effects model (NONMEM) software. In addition, the effect of CYP2C19 polymorphism on proton pump inhibition by lansoprazole was investigated using 24-h intragastric pH monitoring in the 32 subjects. Time course of serum lansoprazole concentration following intravenous short infusion was well described by a 2-compartment model. The mean volume of the central and peripheral compartments was 0.110 and 0.201 l/kg, respectively. The mean inter-compartment clearance was estimated to be 0.0882 l/h/kg. The population mean value of systemic clearance in the homoEM (CYP2C19 1/ 1), heteroEM (CYP2C19 1/2 and 1/3), and PM (CYP2C19 2/2, 2/3, and 3/3) groups was 0.179, 0.109, and 0.038 l/h/kg, respectively. The mean intragastric pH following twice-daily doses of 30 mg lansoprazole was approximately 6, 5, and 4 in the PM, heteroEM, and homoEM groups, respectively. These findings indicate that large interindividual variability exists in the pharmacokinetics of intravenously administered lansoprazole, but that twice-daily infusion of a 30 mg dose leads to significant and sustained proton pump inhibition, even in the homoEM group, despite the short elimination half-life of the drug. [Abstract/Link to Full Text]

Kurokawa J
Compartmentalized regulations of ion channels in the heart.
Biol Pharm Bull. 2007 Dec;30(12):2231-7.
The rate and force of contraction of the heart are precisely controlled by compartmentalized regulation of cardiac ion channels which determine electrical activities. It is known that modulation of cardiac ion channels, which is caused by drug administration, sympathetic nervous system stimulation and gender difference, can increase risks of lethal arrhythmias in carriers of inherited disease mutations. These modulations are thought to also be involved in common cardiac arrhythmias. Because many signaling molecules are localized within single cells, an understanding of the molecular basis of compartmentalized regulation of cardiac channels is a key for understanding and treating the lethal arrhythmias. In this review, I will discuss molecular mechanisms of compartmentalized regulation of cardiac ion channels via drugs, cAMP and sex hormones. [Abstract/Link to Full Text]

Sasaki Y, Nagumo S
Rapid identification of Curcuma longa and C. aromatica by LAMP.
Biol Pharm Bull. 2007 Nov;30(11):2229-30.
We have developed a novel method for the identification of Curcuma longa and C. aromatica called "loop-mediated isothermal amplification (LAMP)," based on trnK gene sequences. LAMP employs four primers that recognize six regions on the target DNA. Cycling elongation was initiated when the four primers were annealed to the target DNA. Amplifications were detected by measuring turbidity due to the formation of magnesium pyrophosphate. We designed allele-specific primer sets for C. longa and C. aromatica, respectively. LAMP using a primer set for C. longa and total DNA extracted from C. longa rhizome (0.5-10.0 ng) as template was detected up to 70 min. On the other hand, in the reaction using a primer set for C. longa and total DNA from C. aromatica as template, no amplifications were detected. The same tendency could be seen in the reactions using a set of primers for C. aromatica. LAMP enabled not only identification but also detection with high specificity. This rapid, specific, sensitive, and convenient method is expected to be applicable to the identification of the botanical origin of commercially available herbal products. [Abstract/Link to Full Text]

Akizawa H, Saito M, Tsukamoto I, Ohkura T, Shimizu T, Kitamura Y, Mifune M, Saito Y, Arano Y, Saji H
Effect of carboxyl-group of D-glutamic acid or gamma-carboxy-D-glutamic acid as N-terminal amino acid of (111)in-diethylenetriaminepentaacetic acid-octreotide on accumulation of radioactivity in kidney.
Biol Pharm Bull. 2007 Nov;30(11):2226-8.
To establish a strategy for developing (111)In-diethylenetriaminepentaacetic acid ((111)In-DTPA)-octreotide, a diagnostic radiopharmaceutical agent for tumors, with reduced non-specific renal radio-accumulation, the compounds having D-glutamic acid (Glu) or gamma-carboxy-D-glutamic acid (carboxy-Glu) as the N-terminal amino acid were examined for in vivo radio-distribution. Compounds carrying Glu and carboxy-Glu containing one and two negative charges, respectively, showed lower renal radio-accumulation than that carrying D-phenylalanine. It was revealed that the introduction of a negative charge reduces the renal radio-accumulation independently from the number of negative charges. The present result can be a clue for the development of (111)In-DTPA-octreotides with reduced the renal radio-accumulation. [Abstract/Link to Full Text]

Nishimura H, Hayashi C, Aiba T, Okamoto I, Miyamoto Y, Nakade S, Takeda K, Kurosaki Y
Application of the correlation of in vitro dissolution behavior and in vivo plasma concentration profile (IVIVC) for soft-gel capsules--a pointless pursuit?
Biol Pharm Bull. 2007 Nov;30(11):2221-5.
Plasma concentration profiles of arundic acid ((R)-(-)-2-propyloctanoic acid), an oil-like medicine, administered as soft-gel capsules in human clinical tests were predicted from the dissolution test data of the soft-gel capsules with different storage terms (short- and long-term stored drugs) by applying the in vitro-in vivo correlation (IVIVC). We established two linear-regression IVIVCs, which were characterized by either the in vitro dissolution behaviors against the pH 8.0 dissolution medium or the pH 6.8 dissolution medium containing 2% sodium dodecyl sulfate (SDS), in this study. Also, the prediction accuracies for the in vivo plasma profiles in humans for these two IVIVCs were compared. Regarding dissolution from the long-term stored capsule in pH 8.0 dissolution medium without surfactant, the prediction accuracies of the in vivo plasma profiles in humans were not satisfactory for the obtained IVIVC. The use of pH 6.8 dissolution medium containing 2% SDS, according to the Japanese guideline, improved the dissolution of the long-term stored capsule. Furthermore, the prediction accuracies for the in vivo plasma profiles in humans for these two IVIVCs were compared. The IVIVC established by the in vitro dissolution data obtained with the dissolution medium containing surfactant more effectively predicted the plasma drug concentration profiles following oral administrations of the soft-gel capsules under both storage conditions. [Abstract/Link to Full Text]

Ohki E, Suzuki M, Aoe T, Ikawa Y, Negishi E, Ueno K
Expression of histamine H4 receptor in synovial cells from rheumatoid arthritic patients.
Biol Pharm Bull. 2007 Nov;30(11):2217-20.
The last of four histamine receptors (HRs), H4 receptor (H4R), was identified in the year 2000. Since that time, the H4R has been implicated in cellular mechanisms related to immune systems, inflammatory processes, and allergic reactions. We have previously reported the expression of H4R mRNA in rheumatoid arthritic (RA) synovial cell cultures by means of the RT-PCR method. The present study demonstrates the expression of H4R at the protein level in RA synovial cell cultures. We also identified that the receptor was expressed in two specific types of cell populations, fibroblast-like and macrophage-like cells from RA synovial tissues. In the culture system, the fifth generation of fibroblast-like synoviocytes showed as strong expression of H4R as in the primary culture. The results provide pragmatic evidence for H4R as a novel target in the development of pharmacotherapeutic agents for RA treatment. Furthermore, the culture system used here will be useful for future studies concerning basic molecular mechanisms underlying the pharmacology of H4R. [Abstract/Link to Full Text]

Bhandari KH, Newa M, Yoon SI, Kim JS, Jang KY, Kim JA, Yoo BK, Woo JS, Lee JH, Kim DD, Choi HG, Yong CS
Evaluation of physicochemical properties, skin permeation and accumulation profiles of ketorolac fatty ester prodrugs.
Biol Pharm Bull. 2007 Nov;30(11):2211-6.
The purpose of this study was to evaluate the physicochemical properties, skin permeation and accumulation profiles of model lipophilic ketorolac fatty ester (esters) prodrugs. Ketorolac linoleate (C18:2), oleate (C18:1) and stearate (C18:0) were evaluated for their solubility, capacity factor, enzymatic hydrolysis, chemical stability, and skin permeation and accumulation profiles using the combination of common permeation enhancing techniques such as the use of supersaturated solution of permeants in the enhancer vehicle, lipophilic receptor solution, enhancer pretreatment of skins, removal of stratum corneum and delipidization of skins etc. Esters were highly lipophilic, chemically stable for the duration of observation, enzymatically unstable in hairless mouse skin/liver homogenates and plasma, and impermeable into the receptor solution. Absence of skin permeation, relative enzymatic stability during permeation and chemical stability of these esters could delineate preliminary possibilities for designing safer topical agents without systemic absorption. [Abstract/Link to Full Text]

Jang DS, Lee GY, Kim YS, Lee YM, Kim CS, Yoo JL, Kim JS
Anthraquinones from the seeds of Cassia tora with inhibitory activity on protein glycation and aldose reductase.
Biol Pharm Bull. 2007 Nov;30(11):2207-10.
Nine anthraquinones, aurantio-obtusin (1), chryso-obtusin (2), obtusin (3), chryso-obtusin-2-O-beta-D-glucoside (4), physcion (5), emodin (6), chrysophanol (7), obtusifolin (8), and obtusifolin-2-O-beta-D-glucoside (9), isolated from an EtOAc-soluble extract of the seeds of Cassia tora, were subjected to in vitro bioassays to evaluate their inhibitory activity against advanced glycation end products (AGEs) formation and rat lens aldose reductase (RLAR). Among the isolates, compounds 6 and 8 exhibited a significant inhibitory activity on AGEs formation with observed IC(50) values of 118 and 28.9 microM, respectively, in an AGEs-bovine serum albumin (BSA) assay by specific fluorescence. Furthermore, compounds 6 and 8 inhibited AGEs-BSA formation more effectively than aminoguanidine, an AGEs inhibitor, by indirect AGEs-ELISA. N(epsilon)-Carboxymethyllysine (CML)-BSA formation was also inhibited by compounds 6 and 8. Whereas compounds 1, 4, and 6 showed a significant inhibitory activity on RLAR with IC(50) values of 13.6, 8.8, and 15.9 microM, respectively. [Abstract/Link to Full Text]

Jin JS, Kakiuchi N, Hattori M
Enantioselective oxidation of enterodiol to enterolactone by human intestinal bacteria.
Biol Pharm Bull. 2007 Nov;30(11):2204-6.
In the course of our experiments on the metabolic conversion of lignans to the estrogenic substances enterodiol (END) and enterolactone (ENL) by human intestinal flora, we isolated two anaerobes, Ruminococcus sp. END-1 and strain END-2, capable of oxidizing END. The former selectively converted (-)-END to (-)-ENL, while the latter selectively converted (+)-END to (+)-ENL, indicating enantioselective oxidation by intestinal bacteria. [Abstract/Link to Full Text]

Han SJ, Bae EA, Trinh HT, Yang JH, Youn UJ, Bae KH, Kim DH
Magnolol and honokiol: inhibitors against mouse passive cutaneous anaphylaxis reaction and scratching behaviors.
Biol Pharm Bull. 2007 Nov;30(11):2201-3.
The antiallergic effects of magnolol and honokiol, isolated from the bark of Magnolia obovata (family Magnoliaceae), were investigated both in vitro and in vivo. Magnolol and honokiol potently inhibited passive cutaneous anaphylaxis reactions in mice induced by IgE-antigen complex as well as compound 48/80-induced scratching behaviors. These constituents exhibited not only potent inhibitory activity on the degranulation of RBL-2H3 cells induced by IgE-antigen complex, with IC(50) values of 45 and 55 muM, respectively, but also inhibited the protein expressions of IL-4 and TNF-alpha. Based on these findings, magnolol and honokiol may improve IgE-induced allergic diseases. [Abstract/Link to Full Text]

Yoon SH, Han EJ, Sung JH, Chung SH
Anti-diabetic effects of compound K versus metformin versus compound K-metformin combination therapy in diabetic db/db mice.
Biol Pharm Bull. 2007 Nov;30(11):2196-200.
Compound K (CK) is a major intestinal metabolite of ginsenosides derived from ginseng radix. In our preliminary studies, CK has shown to exhibit anti-hyperglycemic effect through its insulin-secreting action, similar to that of insulin secretagogue sulfonylureas. Metformin, a biguanide, improves insulin resistance by reducing gluconeogenesis and enhancing peripheral glucose uptake, promoting reduction of the plasma glucose level. The aim of this study was to compare the anti-diabetic effects of CK and metformin due to differences in their mechanisms of action and also to investigate whether treatment of CK and metformin in combination show synergistic or additive effects compared to each drug alone. Seven week-old male db/db mice were treated for 8 weeks. CK was given at a dose of 10 mg/kg, metformin at 150 mg/kg and the same dosage of each drug was applied to CK plus metformin combination group. Significant improvements were observed in plasma glucose and insulin levels, homeostasis model assessment-insulin resistance (HOMA-IR) index and in hematoxylin and eosin-stained liver tissues in combination group. Although further studies to elucidate the benefits of co-administration of CK and metformin are needed, our findings may provide basis to the discovery of a new combination therapy on diabetes control in type 2 diabetics. [Abstract/Link to Full Text]

Lee CK, Park KK, Lim SS, Park JH, Chung WY
Effects of the licorice extract against tumor growth and cisplatin-induced toxicity in a mouse xenograft model of colon cancer.
Biol Pharm Bull. 2007 Nov;30(11):2191-5.
Cisplatin is one of the most effective chemotherapeutic agents and plays a major role in the treatment of a variety of human solid tumors. However, its toxicity limits the clinical use. Recently, the administration of antioxidants has been suggested to protect against cisplatin-induced nephrotoxicity. The present study was designed to estimate the antitumor activity of the licorice extract alone and in combination with cisplatin, and its protective potential against cisplatin-induced toxicity in a mouse xenograft model. The administration of the licorice extract significantly inhibited tumor growth in BALB/C mice inoculated with CT-26 colon cancer cells. The combination of the licorice extract and cisplatin diminished the therapeutic efficacy of cisplatin but promoted considerably antitumor activity of the licorice extract. In mice with cisplatin treatment for 15 d, the serum levels of blood urea nitrogen and creatinine remarkably were increased by kidney damage, and the serum alanine aminotransferase and aspartate aminotransferase levels were elevated by liver damage. The administration of the licorice extract plus cisplatin recovered these functional indices in the kidney and liver to almost the control levels. In addition, the administration of the licorice extract significantly reduced the cisplatin-induced oxidative stress. Taken together, the administration of the licorice extract inhibits the growth of mouse colon carcinoma without any adverse effects, and reduces the cisplatin-induced toxicity. Therefore, the licorice extract may be a candidate for an anticancer and chemopreventive agent. However, cancer patients with cisplatin therapy should avoid the supplementation of the licorice extract. [Abstract/Link to Full Text]

Hirai T, Fukui Y, Motojima K
PPARalpha agonists positively and negatively regulate the expression of several nutrient/drug transporters in mouse small intestine.
Biol Pharm Bull. 2007 Nov;30(11):2185-90.
A systematic analysis to examine the effects of peroxisome proliferator-activated receptor (PPAR)alpha agonists on the expression levels of all the nutrient/drug plasma-membrane transporters in the mouse small intestine was performed. Transporter mRNAs that were induced or repressed by two independent PPARalpha-specific agonists were identified by a genome-wide microarray method, and the changes were confirmed by real-time PCR using RNA isolated from the intestines and livers of wild-type and PPARalpha-null mice. Expression levels of seven nutrient/drug transporters (Abcd3, Octn2/Slc22a5, FATP2/Slc27a2, Slc22a21, Mct13/Slc16a13, Slc23a1 and Bcrp/Abcg2) in the intestine were up-regulated and the expression level of one (Mrp1/Abcc1) was down-regulated by PPARalpha; although the previously report that the H(+)/peptide co-transporter 1 (Pept1) is up-regulated by PPARalpha was not replicated in our study. We propose that the transport processes can be coordinately regulated with intracellular metabolism by nutrient nuclear receptors. [Abstract/Link to Full Text]

Sommani P, Yamashita K, Miyoshi T, Tsunemine H, Kodaki T, Mori H, Hirota K, Arai T, Sasada M, Makino K
Inhibitory effect of 6-formylpterin on HIF-1alpha protein accumulation.
Biol Pharm Bull. 2007 Nov;30(11):2181-4.
Hypoxia-inducible factor-1 (HIF-1) is a main regulator of metabolic adaptation to hypoxia. HIF-1alpha is induced by hypoxia, or by hypoxia-mimicking reagents, such as desferrioxamine (DFX), under a normoxic condition. A xanthine oxidase inhibitor, 6-formylpterin (6FP), is reported to exert its functions on reactive oxygen species (ROS) modulation. In this study, we investigated the effect of 6FP on HIF-1alpha expression under a DFX-treated or hypoxic condition. 6FP decreased HIF-1alpha expression at the protein level, but not at the mRNA level, in a dose-dependent manner, and this suppressive effect was reversed by the antioxidant, N-acetyl-L-cysteine (NAC). Furthermore, the ROS generated by 6FP was reversed with NAC coincubation. These findings suggest that intracellular ROS generated by 6FP decreased the HIF-1alpha protein accumulation under a DFX-treated or hypoxic condition. [Abstract/Link to Full Text]

Hirano Y, Tsunoda M, Shimosawa T, Fujita T, Funatsu T
Measurement of catechol-O-methyltransferase activity in the brain of Dahl salt-sensitive rats.
Biol Pharm Bull. 2007 Nov;30(11):2178-80.
Metabolism by catechol-O-methyltransferase (COMT) is one of the inactivation pathways of catecholamines (CAs), which are important hormones in regulating blood pressure both in central and in peripheral sympathetic nerve endings. We have reported the rapid determination method of COMT activity using high-performance liquid chromatography (HPLC)-fluorescence detection. In the present study, we applied the method to brain tissues, cerebral cortex, cerebellum, hypophysis and hypothalamus. COMT activities were assessed by measuring normetanephrine with the use of norepinephrine as an endogenous substrate. We examined the COMT activities in the brains of Dahl salt-sensitive (DS) rats given normal-salt or high-salt diet for 13 weeks, and found that membrane-bound COMT activities in the cerebral cortex were significantly reduced in high-salt loaded DS rats compared with normal-salt loaded DS rats. [Abstract/Link to Full Text]

Kokubun H, Fukawa M, Matoba M, Hoka S, Yamada Y, Yago K
Pharmacokinetics and variation in the clearance of oxycodone and hydrocotarnine in patients with cancer pain.
Biol Pharm Bull. 2007 Nov;30(11):2173-7.
Compound injections of oxycodone and hydrocotarnine are currently used as one of the treatment options for some cases with cancer pain. However, there have been no reports examining the factors that influence oxycodone and hydrocotarnine clearance, so detailed examination is necessary. As for hydrocotarnine, there have been no reports examining the pharmacokinetics. Therefore in this study, we determined the pharmacokinetics of oxycodone and hydrocotarnine in patients with cancer pain. The study was conducted on 19 patients, in whom pain control was attempted by using the compound injections of oxycodone and hydrocotarnine. We used HPLC-electrochemical detector (ECD) to determine oxycodone and hydrocotarnine serum concentrations, and used the nonlinear least-squares method (MULTI) for calculation of the pharmacokinetic parameters. Furthermore, we examined the factors that influence the clearance of oxycodone and hydrocotarnine by multiple regression analysis (step wise method). The pharmacokinetic parameters were as follows: Oxycodone; V(d)=226.7+/-105.5 l (mean+/-S.D.), CL=37.9+/-25.1 l/h, t(1/2)=4.1+/-1.9 h. Hydrocotarnine; V(d)=276.8+/-237.2 l, CL=95.1+/-64.3 l/h, t(1/2)=2.0+/-0.7 h. The clearance of oxycodone represented by a regression formula was significantly correlated to the age, the presence or absence of within 7 d on the death or liver metastasis, or of the heart failure of the patients. The clearance of hydrocotarnine represented by a regression formula was significantly correlated to the presence or absence of within 7 d on the death or liver metastasis, or of the heart failure of the patients. The clearance also indicated that oxycodone concentration in the blood was likely to be higher in patients having these factors. Oxycodone/hydrocotarnine compound injections should be used with caution and dose reduction may be necessary in such populations. [Abstract/Link to Full Text]

Takaai M, Suzuki H, Ishida K, Tahara K, Hashimoto Y
Pharmacokinetic analysis of transcellular transport of levofloxacin across LLC-PK1 and Caco-2 cell monolayers.
Biol Pharm Bull. 2007 Nov;30(11):2167-72.
To characterize the membrane transport responsible for the renal excretion and intestinal absorption of levofloxacin, we performed pharmacokinetic analysis of transcellular transport across LLC-PK(1) and Caco-2 cell monolayers. Transcellular transport of levofloxacin in LLC-PK(1) cells was greater in the basolateral-to-apical direction than in the opposite direction. Pharmacokinetic analysis indicated that basolateral uptake was the direction-determining step for the transcellular transport of levofloxacin in LLC-PK(1) cells. The apical efflux clearance of levofloxacin in LLC-PK(1) cells was increased at the medium pH 6 as compared with at pH 8, suggesting that membrane transport characteristics of levofloxacin are apparently similar to those of a prototypical organic cation, tetraethylammonium. On the other hand, transcellular transport of levofloxacin in Caco-2 cells was only slightly greater in the basolateral-to-apical direction than in the opposite direction. The apical efflux clearance of levofloxacin in Caco-2 cells was greater than basolateral efflux clearance, and apical influx clearance was greater than any other membrane transport clearance. In addition, the apical uptake of levofloxacin as well as quinidine in Caco-2 cells was inhibited significantly by nicotine and imipramine. The findings indicated that some transporters are responsible not only for the efflux but also for the influx of levofloxacin at the apical membrane of Caco-2 cells. [Abstract/Link to Full Text]

Sunada Y, Nakamura S, Kamei C
Effects of Lactobacillus acidophilus strain L-55 on experimental allergic rhinitis in BALB/c mice.
Biol Pharm Bull. 2007 Nov;30(11):2163-6.
We investigated the effect of Lactobacillus acidophilus strain L-55 isolated from infant feces on experimental allergic rhinitis in BALB/c mice. The heat-treated cells of strain L-55 were orally administrated for 4 consecutive weeks to mice sensitized by ovalbumin (OVA), and nasal symptoms (sneezing and nasal rubbing) induced by OVA challenge were evaluated. Strain L-55 at doses of 1 and 10 mg cells/mouse significantly inhibited nasal symptoms by repeated administration over a period of 2 weeks. Furthermore, we measured the level of OVA-specific IgE titers in the serum by passive cutaneous anaphylaxis (PCA) reaction. PCA titers in the sera from mice administrated strain L-55 were significantly lowered compared with the control. These results suggest that oral administration of strain L-55 may be useful for alleviating the nasal symptoms of allergic rhinitis. [Abstract/Link to Full Text]

Ishida K, Kayano Y, Taguchi M, Hashimoto Y
Simulation for clinical repeated-dose pharmacokinetic trials applying a peak-and-trough sampling design to estimate oral clearance.
Biol Pharm Bull. 2007 Nov;30(11):2159-62.
We performed a simulation for the clinical pharmacokinetic study, in which blood was sampled at two time points corresponding to the peak concentration (C(peak)) and trough concentration (C(trough)) following repetitive oral drug administration to subjects. We estimated the approximate oral clearance (CL/F(approx)) as 2.D/(C(peak).tau+C(trough).tau), where D is the dose, and tau is the dosing interval. The CL/F(approx) value was accurate for drugs with a long-elimination half-life, and the estimation error of the CL/F value was slightly increased for drugs with a shorter elimination half-life. The accuracy of CL/F(approx) in each subject was not affected by the magnitude of the interindividual pharmacokinetic variability, but was significantly decreased by the larger measurement error of drug concentrations (or intraindividual pharmacokinetic variability). We further performed several computer simulations to mimic statistical hypothesis testing following the clinical repeated-dose pharmacokinetic trials. The statistical power to detect the difference of oral clearance between two groups was marginally dependent on the measurement error of drug concentration, but was highly dependent on the interindividual pharmacokinetic variability. These findings suggested that the peak-and-trough sampling design to estimate the CL/F(approx) value is useful for clinical repeated-dose pharmacokinetic trials, and that the study design and protocol should be evaluated carefully by computer simulation prior to a real clinical trial. [Abstract/Link to Full Text]

Koue T, Kubo M, Funaki T, Fukuda T, Azuma J, Takaai M, Kayano Y, Hashimoto Y
Nonlinear mixed effects model analysis of the pharmacokinetics of aripiprazole in healthy Japanese males.
Biol Pharm Bull. 2007 Nov;30(11):2154-8.
The population pharmacokinetic parameters of aripiprazole in healthy Japanese males were estimated using a nonlinear mixed effects model (NONMEM) program. Pharmacokinetic data for population analysis were obtained from the single-dose (24 subjects), multiple-dose (15 subjects), and itraconazole-coadministration (27 subjects) trials. The time course of plasma aripiprazole concentration following oral administration was well described by a two-compartment model with first-order input. The mean values of the absorption lag time (ALAG) and absorption rate constant (KA) were estimated to be 0.805 h and 2.65 h(-1), respectively. The mean volume of the central (V(1)/F) and peripheral (V(2)/F) compartment was 3.84 and 1.54 l/kg, respectively, and the mean value of inter-compartment clearance (Q/F) was 0.168 l/h/kg. Oral clearance (CL/F) was estimated to be 0.0645 l/h/kg in the group with CYP2D6*1/*1, *1/*2 and *2/*2. The decrease in CL/F was estimated to be 0.0135 l/h/kg in the group with CYP2D6*1/*5, *1/*10, *2/*5, *2/*10, and *2/*41, and 0.0293 l/h/kg in the group with CYP2D6*5/*10, *10/*10, and *41/*41. The plasma concentration of aripiprazole was increased by coadministration of itraconazole, and the decrease in CL/F was estimated to be 0.0181 l/h/kg. [Abstract/Link to Full Text]

Takekuma Y, Takenaka T, Yamazaki K, Ueno K, Sugawara M
Stereoselective metabolism of racemic carvedilol by UGT1A1 and UGT2B7, and effects of mutation of these enzymes on glucuronidation activity.
Biol Pharm Bull. 2007 Nov;30(11):2146-53.
Carvedilol, an alpha- and beta-adrenergic blocking drug, is mainly metabolized by CYP2D6, UGT1A1, UGT2B4 and UGT2B7. This drug is administered orally as a racemic mixture of R(+)- and S(-)-enantiomers. It has been reported that CYP2D6 prefers metabolizing S-carvedilol to R-carvedilol stereoselectively. On the other hand, stereoselective metabolism of carvedilol by UGTs is still unclear. Moreover, we have reported that patients with chronic heart failure who had polymorphism in CYP2D6, UGT1A1 and/or UGT2B7 had lower metabolic activity and oral clearance than did patients with no polymorphism. The aim of this study was to clarify stereoselective metabolism of carvedilol by UGT1A1 and UGT2B7 and to determine by using a recombinant enzyme-introduced mutation whether genetic mutation in UGT1A1 and UGT2B7 causes reduction in metabolic activity for carvedilol. A glucuronidation assay using human liver microsomes and recombinant UGT1A1 and UGT2B7 expressed in HeLa cells demonstrated that UGT1A1 prefers metabolizing R-carvedilol to S-carvedilol. On the other hand, UGT2B7 prefers metabolizing S-carvedilol to R-carvedilol. Moreover, G71R mutation of UGT1A1 reduced both affinity and capacity but did not affect stereoselective metabolism. On the other hand, both A71S and H268Y mutations of UGT2B7 reduced capacity but did not affect affinity and, as a result, the efficiency of metabolism was remarkably reduced. However, as in the case of UGT1A1, neither of the mutations affected stereoselective metabolism. [Abstract/Link to Full Text]

Yoon JH, Shim JS, Cho Y, Baek NI, Lee CW, Kim HS, Hwang JK
Depigmentation of melanocytes by isopanduratin A and 4-hydroxypanduratin A isolated from Kaempferia pandurata ROXB.
Biol Pharm Bull. 2007 Nov;30(11):2141-5.
This study was carried out to investigate the in vitro effects of isopanduratin A and 4-hydroxypanduratin A isolated from Kaempferia pandurata ROXB. on melanin biosynthesis and tyrosinase activity. Two chalcone compounds, isopanduratin A and 4-hydroxypanduratin A, were isolated from the ethyl acetate fraction of ethanol extract as the active principles. Compared with phenylthiourea (IC(50)=34.3 microM) as a positive control, the depigmentation IC(50) values for isopanduratin A and 4-hydroxypanduratin A were 10.6 microM and 23.2 microM, respectively. The compounds also significantly inhibited the activity of tyrosinase, the enzyme that converts DOPA (3,4-dihydroxyphenylalanine) to dopachrome in the biosynthetic process of melanin. The IC(50) values of isopanduratin A and 4-hydroxypanduratin A for tyrosinase were 10.5 microM and >30 microM, respectively, while that of phenylthiourea was 47.6 microM. The tyrosinase protein level was also significantly decreased by isopanduratin A and 4-hydroxypanduratin A. The results indicate that isopanduratin A and 4-hydroxypanduratin A isolated from K. pandurata ROXB. are promising compounds that could be useful for treating hyperpigmentation as skin-whitening agents. [Abstract/Link to Full Text]

Hasegawa T, Kawazome A, Yanagimoto G, Hayashi T, Seki T, Akimoto M, Todo H, Sugibayashi K
Analysis of skin disposition of flurbiprofen after topical application using dual agar gel discs-inserted rats.
Biol Pharm Bull. 2007 Nov;30(11):2135-40.
Most of the drug fraction penetrating the skin after topical application is taken up by the cutaneous blood flow, although the rest directly migrates into deeper tissues such as the subcutis and muscle. A new in situ experimental hairless rat model was designed to distinguish these fractions of topically applied drugs. Flurbiprofen, a non-steroidal anti-inflammatory drug, was selected as the model drug. In this model, two agar gel discs were subcutaneously inserted into the abdominal region of hairless rats as a drug receptor, and a topical formulation containing the drug was placed above either side of the gel disc. Plasma and agar levels of flurbiprofen were followed every 2 h over 10 h. The migration fraction of the drug into the systemic circulation and that directly to subcutaneous tissues were calculated to be 99.8% and 0.2% against the total amount which penetrated the skin, and the drug ratios into agar gel from the systemic circulation and not from the systemic circulation (i.e. directly migrated from the formulation) were 16.0% and 84.0%, respectively, at 10 h. This in situ drug disposition profile in skin was similar to the in vivo profile calculated from the in vivo muscle amount of flurbiprofen using muscle clearance. These results clearly suggest that the present in situ experimental model is a valuable tool for easy analysis of the skin disposition of topically applied drugs. [Abstract/Link to Full Text]

Lee JH, Choi SH, Kwon OS, Shin TJ, Lee JH, Lee BH, Yoon IS, Pyo MK, Rhim H, Lim YH, Shim YH, Ahn JY, Kim HC, Chitwood DJ, Lee SM, Nah SY
Effects of ginsenosides, active ingredients of Panax ginseng, on development, growth, and life span of Caenorhabditis elegans.
Biol Pharm Bull. 2007 Nov;30(11):2126-34.
The backbone structure of ginsenosides, active ingredients of Panax ginseng, is similar with that of sterol, especially cholesterol. Caenorhabditis elegans (C. elegans) is one of free living nematodes and is well-established animal model for biochemical and genetic studies. C. elegans cannot synthesize de novo cholesterol, although cholesterol is essential requirement for its growth and development. In the present study, we investigated the effects of ginseng total saponins (GTS) on the average brood size, growth, development, worm size, and life span of C. elegans in cholesterol-deprived and -fed medium. Cholesterol deprivation caused damages on normal growth, reproduction, and life span of worms throughout F1 to F3 generations. GTS supplement to cholesterol-deprived medium restored the growth, reproduction, and life span of worms as much as cholesterol alone-fed medium. GTS co-supplement to cholesterol-fed medium not only promoted worm reproduction but also induced bigger worms and faster growth than cholesterol-fed medium. In study to identify which ginsenosides are responsible for life span restoring effects of GTS, we found that ginsenoside Rc supplement not only restored life span of worms grown in cholesterol-deprived medium but also prolonged life span of worms grown in cholesterol-fed medium. Worms grown in medium supplemented with ginsenoside Rb(1) or Rc to cholesterol-deprived medium exhibited strong filipin staining, in which filipin forms tight and specific complexes with 3beta-hydroxy sterols. These results show a possibility that ginsenosides could be utilized by C. elegans as a sterol substitute and further indicate that ginsenoside Rc is the component of Panax ginseng that prolongs the life span of C. elegans. [Abstract/Link to Full Text]

Kim SH, Shin EJ, Kim ED, Bayaraa T, Frost SC, Hyun CK
Berberine activates GLUT1-mediated glucose uptake in 3T3-L1 adipocytes.
Biol Pharm Bull. 2007 Nov;30(11):2120-5.
It has recently been known that berberine, an alkaloid of medicinal plants, has anti-hyperglycemic effects. To explore the mechanism underlying this effect, we used 3T3-L1 adipocytes for analyzing the signaling pathways that contribute to glucose transport. Treatment of berberine to 3T3-L1 adipocytes for 6 h enhanced basal glucose uptake both in normal and in insulin-resistant state, but the insulin-stimulated glucose uptake was not augmented significantly. Inhibition of phosphatidylinositol 3-kinase (PI 3-K) by wortmannin did not affect the berberine effect on basal glucose uptake. Berberine did not augment tyrosine phosphorylation of insulin receptor (IR) and insulin receptor substrate (IRS)-1. Further, berberine had no effect on the activity of the insulin-sensitive downstream kinase, atypical protein kinase C (PKCzeta/lambda). However, interestingly, extracellular signal-regulated kinases (ERKs), which have been known to be responsible for the expression of glucose transporter (GLUT)1, were significantly activated in berberine-treated 3T3-L1 cells. As expected, the level of GLUT1 protein was increased both in normal and insulin-resistant cells in response to berberine. But berberine affected the expression of GLUT4 neither in normal nor in insulin-resistant cells. In addition, berberine treatment increased AMP-activated protein kinase (AMPK) activity in 3T3-L1 cells, which has been reported to be associated with GLUT1-mediated glucose uptake. Together, we concluded that berberine increases glucose transport activity of 3T3-L1 adipocytes by enhancing GLUT1 expression and also stimulates the GLUT1-mediated glucose uptake by activating GLUT1, a result of AMPK stimulation. [Abstract/Link to Full Text]

Jin JS, Zhao YF, Nakamura N, Akao T, Kakiuchi N, Min BS, Hattori M
Enantioselective dehydroxylation of enterodiol and enterolactone precursors by human intestinal bacteria.
Biol Pharm Bull. 2007 Nov;30(11):2113-9.
During the course of experiments on the transformation of lignans to phytoestrogenic substances, such as enterodiol (END) and enterolactone (ENL), a previously isolated bacterium, Eubacterium (E.) sp. strain SDG-2, capable of phenolic p-dehydroxylation in the biotransformation of secoisolariciresinol diglucoside to END and ENL, was concluded to be Eggerthella (Eg.) lenta (Eg. sp. SDG-2) on the basis of 16S rRNA gene sequence analysis. The bacterium could transform (+)-dihydroxyenterodiol (DHEND, 3a) to (+)-END (1a), but not for (-)-DHEND (3b) to (-)-END (1b) under anaerobic conditions. By incubation of a mixture of (+)- and (-)-dihydroxyenterolactone (DHENL, 4a and 4b) with Eg. sp. SDG-2, only (-)-DHENL (4b) was converted to (-)-ENL (2b), selectively. On the other hand, we isolated a different bacterium, strain ARC-1, capable of dehydroxylating (-)-DHEND (3b) to (-)-END (1b) from human feces. Strain ARC-1 could transform not only (-)-DHEND (3b) to (-)-END (1b), but also (+)-DHENL (4a) to (+)-ENL (2b). However, the bacterium could not transform (+)-DHEND (3a) and (-)-DHENL (4b). Both bacterial strains demonstrated different enantioselective dehydroxylation. [Abstract/Link to Full Text]

Cho JY
Effect of L-cycloserine on cellular responses mediated by macrophages and T cells.
Biol Pharm Bull. 2007 Nov;30(11):2105-12.
In this study, we examined the immunoregulatory roles of L-cycloserine (L-CS), a sphingolipid metabolism regulator with inhibitory activity of serine palmitoyltransferase (SPT), on immune responses mediated by monocytes/macrophages and T cells. Mitogenic responses of splenic lymphocytes induced by LPS, PHA, and Con A were very strongly suppressed by L-CS with IC(50) values ranging from 0.5 to 1 muM. In contrast, this compound less strongly blocked IL-2-induced CD8+ CTLL-2 cell proliferation with an IC(50) value of 540 muM. Interestingly, L-CS enhanced the number of IL-4-producing helper T cells, indicating the favored induction of Th2 condition. Although tumor necrosis factor (TNF)-alpha and nitric oxide (NO) production was not altered under 10% FCS condition, U937 cell-cell adhesion as well as the surface level of adhesion molecules (CD29 and CD98) were significantly suppressed by L-CS. In particular, reduced serum level (5%) under L-CS treatment strongly enhanced the production of TNF-alpha and the inhibitory potency of NO production and cell adhesion. Finally, sphingolipids (D-sphingosine and DL-dihydrosphingosine) did not remarkably abrogate L-CS-mediated T cell proliferation. Therefore our data suggest that de novo sphingolipid metabolism may represent an important aspect of immunomodulatory activities mediated by T cells and macrophages/monocytes, depending on serum level. [Abstract/Link to Full Text]

Rocha ML, Bendhack LM
Effects of K+ channel modulators on oscillatory contractions in sinoaortic denervated rat aortas.
Biol Pharm Bull. 2007 Nov;30(11):2098-104.
Sinoaortic denervated (SAD) rats present arterial pressure lability without sustained hypertension. We investigated the relation between sinoaortic denervation and the occurrence of oscillatory contractions in SAD rat aortas, as well as the effect of various K(+) channel modulators on these oscillations. Aortas were removed and concentration-effect curves to phenylephrine (0.01 to 10 muM) were constructed in arteries from SAD and Sham-operated rats in order to verify the occurrence of oscillations. We also evaluated the effects of various K(+) channel modulators on these oscillations. Only SAD rat aortas exhibited oscillatory contractions. Tetraethylammonium increased the frequency (28.5+/-3.5 to 41.5+/-4.5 counts/5 min) and amplitude (0.435+/-0.07 to 0.630+/-0.09 g) of the oscillations. Apamin and 4-aminopyridine did not alter the oscillations. Barium chloride converted the oscillatory contractions to a tonic contraction. Pinacidil rapidly blocked the oscillatory contractions and glibenclamide evoked reduction in amplitude from 0.410+/-0.07 to 0.180+/-0.06 g. Iberiotoxin increased the frequency of oscillatory contractions (from 28.0+/-3.5 to 51.5+/-7.5 counts/5 min) but decreased the amplitude (from 0.410+/-0.08 to 0.195+/-0.2 g). Our results demonstrate that SAD rat aortas exhibit oscillatory contractions and K(+) channels, mainly K(ATP) and BK(Ca), play a dominant role in these oscillations. [Abstract/Link to Full Text]

Maruyama M, Matsunaga T, Harada E, Ohmori S
Comparison of basal gene expression and induction of CYP3As in HepG2 and human fetal liver cells.
Biol Pharm Bull. 2007 Nov;30(11):2091-7.
Human fetal liver (HFL) cell culture was initiated from a pool of six normal human liver tissues. The proliferation and viability of HFL cells were evaluated using 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide assay, and the cells increased by more than 100-fold by culture for 15 d. The levels of expression of albumin (ALB), hepatocyte nuclear factor 4alpha, hepatocyte growth factor, CYP3A4, CYP3A5, and CYP3A7 mRNAs in HFL cells increased with culture period, while that of alpha-fetoprotein (AFP) mRNA decreased gradually. In HepG2 cells, however, the expression levels of ALB and AFP mRNAs were not changed, and the levels of expression of CYP3A4, CYP3A5, and CYP3A7 mRNAs decreased gradually. The mRNA expression of major CYP isoforms including CYP3As, i.e., CYP1A2, CYP2A6, CYP2B6, CYP2C (2C9 and 2C19), CYP2D6, and CYP2E1, could be detected in HepG2 cells. With the exception of CYP1A2, all of the CYP mRNAs expressed in HepG2 cells were detected in HFL cells. In HFL cells, CYP3A4 and CYP3A7 mRNA expression levels were markedly up-regulated by dexamethasone (DEX), but not by rifampicin (RIF). CYP3A5 mRNA expression was increased to a level 3-fold greater than control by DEX. On the other hand, CYP3A4, CYP3A5, and CYP3A7 mRNA expression levels in HepG2 cells were increased from 2- to 3-fold by treatment with DEX and RIF. Pregnane X receptor mRNA was expressed in HepG2 cells, but not HFL cells. These results indicate that the character of HFL cells with regard to CYP expression was different from that of HepG2 cells. [Abstract/Link to Full Text]

Tokunaga S, Takeda Y, Shinomiya K, Yamamoto W, Utsu Y, Toide K, Kamei C
Changes of sleep patterns in rats with chronic constriction injury under aversive conditions.
Biol Pharm Bull. 2007 Nov;30(11):2088-90.
In the present study, we investigated the changes of sleep parameters in rats with chronic constriction injury (CCI) under aversive conditions. The electroencephalogram (EEG) in the frontal cortex of CCI rats and electromyogram (EMG) were measured over 6 h by placing rats on sandpaper as an aversive condition, to compare with rats placed on sawdust. Six days after CCI surgery, the rats exhibited significant mechanical allodynia, and also had neuropathic pain. When rats were placed on sawdust, no significant difference was observed between the CCI group and sham-operated control group in sleep latency, total waking time, total non-REM sleep time and total REM sleep time. On the other hand, when CCI rats were placed on sandpaper, a significant increase was observed in sleep latency and total waking time compared with the sham group; however, no significant difference was observed in the total non-REM sleep time and total REM sleep time between these two groups. These results indicate that an important factor of sleep disturbance in CCI rats is not only damage to the nerves but also being under aversive conditions. In addition, it was found that CCI rats placed on sandpaper as an aversive condition can serve as a new sleep disturbance model. [Abstract/Link to Full Text]

Recent Articles in Chemical & Pharmaceutical Bulletin (Tokyo)

Maruyama T, Sugimoto N, Kuroyanagi M, Kim IH, Kamakura H, Kawasaki T, Fujita M, Shimada H, Yamamoto Y, Tada A, Yamazaki T, Goda Y
Authentication and chemical study of isodonis herba and isodonis extracts.
Chem Pharm Bull (Tokyo). 2007 Nov;55(11):1626-30.
Isodonis Herba is used as a Japanese dietary supplement and folk medicine. The extract of the herb (Isodonis extract) is also used as a food additive whose major compound is enmein (1). Here we compared internal transcribed spacer sequences of nuclear ribosomal DNA from Isodonis Herba available on the Japanese and Chinese crude drug markets, and found that the former derived from Isodon japonicus and Isodon trichocarpus, while the latter derived from distinct species such as Isodon eriocalyx. The liquid chromatography/mass spectrometry profiles of Isodonis Herba were classified into four chemotypes (A to D) according to the ratio of the major constituents. Types B and C contained 1 and oridonin (2) as major components, respectively. An intermediate (or mixed) form of types B and C in various ratios was designed type A. Type D contained eriocalyxin B (3) as its major component. Japanese herba were types A-C, while Chinese herba were types C and D. The commercial Isodonis extract products tested were classified as type D, suggesting that they originated from Chinese Herba. Understanding the relationship between extract constituents and DNA profiles is important for the official specification of dietary supplements and food additives of plant origin. [Abstract/Link to Full Text]

Hashizaki K, Kageyama T, Inoue M, Taguchi H, Ueda H, Saito Y
Study on preparation and formation mechanism of n-alkanol/water emulsion using alpha-cyclodextrin.
Chem Pharm Bull (Tokyo). 2007 Nov;55(11):1620-5.
Surfactants are usually used for the preparation of emulsions; however, some have an adverse effect on the human body such as skin irritation, hemolysis, and protein denaturation, etc. In this study, we examined the preparation and formation mechanism of n-alkanol/water emulsions using alpha-cyclodextrin (alpha-CD) as an emulsifier. Emulsions were prepared by mixing oil and water phases for 4 min at 2500 rpm using a vortex mixer. The mechanism of emulsification was investigated with some physico-chemical techniques. From phase diagrams of n-alkanol/alpha-CD/water systems, the emulsion phase extended as the chain length of n-alkanols and the amount of alpha-CD added increased. Furthermore, the emulsion was not formed in the region where the n-alkanol/alpha-CD complex didn't precipitate; however, the emulsion was formed in the region where the complex precipitated. In addition, it was clear that the emulsions have a yield stress value and correspond to the Maxwell model from rheological measurement. Our experiments clearly showed that the stable emulsions are formed because the precipitated complexes form a dense film at the oil-water interface and prevent aggregation among dispersed phases. Furthermore, it is suggested that the creation of a three-dimensional network structure formed by precipitated complexes in the continuous phase contributes to the stabilization of the emulsion. Thus, we concluded that the n-alkanol/water emulsions using alpha-cyclodextrin were a kind of the Pickering emulsion. [Abstract/Link to Full Text]

Raghavendra NM, Thampi P, Gurubasavarajaswamy PM, Sriram D
Synthesis and antimicrobial activities of some novel substituted 2-imidazolyl-N-(4-oxo-quinazolin-3(4H)-yl)-acetamides.
Chem Pharm Bull (Tokyo). 2007 Nov;55(11):1615-9.
Several substituted-quinazolin-3(4H)-ones 8-11ad were synthesized by condensation of 2-chloro-N-(4-oxo-substituted-quinazolin-3(4H)-yl)-acetamides with various substituted imidazoles through one pot reaction. Elemental analysis, IR, (1)H-NMR and mass spectral data confirmed the structure of the newly synthesized compounds. Synthesized quinazolin-4-one derivatives were investigated for their antitubercular, antibacterial and antifungal activities. Some of the tested compounds showed good antitubercular activity. None of the synthesized compounds showed significant antibacterial and antifungal activity. [Abstract/Link to Full Text]

Iwaki Y, Akita H
Concise syntheses of cystothiazoles A, C, D, and melithiazol B.
Chem Pharm Bull (Tokyo). 2007 Nov;55(11):1610-4.
A convergent synthesis of cystothiazoles C 1 and D 3 was achieved based on Julia coupling between the functionalized aldehyde 5b, corresponding to left half of the final molecule, and aryl sulfone 6 or 7, bearing a bithiazole moiety, corresponding to right half. Methylation of 1 and 3 gave cystothiazole A 2 and melithiazol B 4, respectively. The overall yield (5 steps from (2R,3S)-3-methylpent-4-yne-1,2-diol 10; 57%) of 5b via the present route was improved in comparison to that of the previously reported functionalized aldehyde 5a (7 steps from 10; 13%). By applying the modified Julia coupling method, selectivity (6E/6Z=20 : 1-26 : 1) toward the (6E)-form of the coupled products (15 or 19) against the corresponding (6Z)-form was improved in comparison to the Wittig method (6E/6Z=4 : 1-6.9 : 1). [Abstract/Link to Full Text]

Nakazaki A, Era T, Kobayashi S
Total synthesis of (+/-)-gleenol and (+/-)-axenol via a functionalized spiro[4.5]decane.
Chem Pharm Bull (Tokyo). 2007 Nov;55(11):1606-9.
Total synthesis of the biologically important axane sesquiterpenes, gleenol (1) and axenol (2), was accomplished through a readily available spiro[4.5]decane. The key features of the synthesis of 1 and 2 include Claisen rearrangement to afford the multi-functionalized spiro[4.5]decane 4 as a single diastereomer in excellent yield, installation of the C7 isopropyl group via ketene dithioacetal instead of direct alkylation and a diastereoselective reduction of ketone under the Birch conditions. [Abstract/Link to Full Text]

Otsuka H, Shitamoto J, He DH, Matsunami K, Shinzato T, Aramoto M, Takeda Y, Kanchanapoom T
Tricalysiosides P-U: Ent-kaurane glucosides and a labdane glucoside from leaves of Tricalysia dubia OHWI.
Chem Pharm Bull (Tokyo). 2007 Nov;55(11):1600-5.
Further extensive isolation work on the 1-BuOH-soluble fraction of a MeOH extract of Tricalysia dubia afforded five new ent-kaurane glucosides (4-8) and one new labdane glucoside (9), together with a known megastigmane glucoside, sammangaoside B (1), and monoterpene glucosides (2, 3). The structures of the new compounds were elucidated by analyses of one- and two-dimensional NMR spectroscopic data. The absolute configuration of the 9-position of sammangaoside B was revised to S and its total stereochemistry was established by the modified Mosher's method. [Abstract/Link to Full Text]

Lan YH, Wang HY, Wu CC, Chen SL, Chang CL, Chang FR, Wu YC
New constituents from stems of Artabotrys uncinatus.
Chem Pharm Bull (Tokyo). 2007 Nov;55(11):1597-9.
Two new compounds, 4,5-dioxoartacinatine (1) and 24-methylenelanosta-7,9(11)-diene-3-one (2), together with thirty known compounds were isolated and characterized from the stems of Artabotrys uncinatus. Structures of the new compounds were determined by spectral analysis. [Abstract/Link to Full Text]

Sato K, Takiguchi Y, Yoshizawa Y, Iwase K, Shimizu Y, Tarui A, Omote M, Kumadaki I, Ando A
Formation of quaternary carbon center with a trifluoromethyl group using a Pd-catalyzed allylation reaction.
Chem Pharm Bull (Tokyo). 2007 Nov;55(11):1593-6.
Synthesis of compounds with quaternary carbons is one of the most attractive reactions in the synthetic chemistry. However, there are only a few reports on synthesis of the compounds with a fluoroalkyl group at a quaternary carbon center. Recently, we reported the synthesis of alpha-trifluoromethylated ketones by the reaction of alpha,beta-unsaturated ketones with CF(3)-I using a Rh catalyst. When the alpha-trifluoromethylated ketones and allyl carbonates were treated with a Pd catalyst, the allylation reaction proceeded smoothly at the trifluoromethylated carbon to give the desired compounds with a trifluoromethylated quaternary carbon center in good to excellent yields. [Abstract/Link to Full Text]

Kobayashi S, Atuchi N, Wakamatsu H, Hattori M, Kawada A, Asano K
Diastereomer-specific effects of double-stranded peptides conjugated with -L-Tyr-L-Phe- or -L-Tyr-D-Phe- residues on tyrosine phosphorylation and inhibition of src(ts)NRK, A431, MCF-7, and DU145 cell growth.
Chem Pharm Bull (Tokyo). 2007 Nov;55(11):1585-92.
The interaction between growth inhibition and chirality, especially of diastereomers, has an important modifying effect on cancer cell proliferation. Previously, we have reported on the design, synthesis, and chemical properties of a series of novel, double-stranded peptides, (y-AA-x-AA)(2)-(CH(2))(12), with -y-AA-x-AA- and -z-AA-y-AA-x-AA- sequences conjugated to the spacer. Here, we extend those results by showing that (D-, L-) and (L-, D-) diastereomers are more potent inhibitors of tyrosine phosphorylation than (L-, L-). Although the replacement of the L-Phe-L-Phe sequence with L-Tyr-L-Phe produces a less active inhibitor, the double-stranded peptide conjugated with L-Tyr-D-Phe is more active than that conjugated with L-Tyr-L-Phe. In addition, we show that SDS-PAGE gel profiles of tyrosine phosphorylation following treatment with bis(y-Tyr-x-Phe)-N,N-dodecane-1,12-diamine appear very similar to profiles of tyrosine phosphorylation following treatment with an analog of the tyrosine kinase inhibitor, erbstatin. Moreover, the level of autophosphorylation of the epidermal growth factor receptor kinase domain (EGFRKD) treated with bis(L-Tyr-D-Phe)-N,N-dodecane-1,12-diamine was lower than that seen following treatment with bis(L-Phe-D-Phe)-N,N-dodecane-1,12-diamine. These data provide new insights for the control of cancer cell proliferation through drug designs which replace the less active -L-Phe-L-Phe- (and -D-Phe-L-Phe-) with the more active -L-Tyr-L-Phe- (and -L-Tyr-D-Phe-) sequence. [Abstract/Link to Full Text]

Mukai J, Tokuyama E, Ishizaka T, Okada S, Uchida T
Inhibitory effect of aroma on the bitterness of branched-chain amino acid solutions.
Chem Pharm Bull (Tokyo). 2007 Nov;55(11):1581-4.
Nutritional products for patients with liver failure available on the Japanese market contain many branched-chain amino acids (BCAAs) such as L-leucine, L-isoleucine, and L-valine, which not only have a bitter taste but also strong, unpleasant odours, leading to low palatability. The palatability of these nutritional products can be significantly improved by the addition of flavoured powders containing various kinds of tastants (sucrose, citric acid, etc.) and odourants (fruit, coffee aromas, etc.). The specific effects of the aroma of flavoured powders have not yet been clearly evaluated. In the present article, the inhibitory effect of aroma on the bitterness of BCAA solutions was examined. The bitterness intensity of a BCAA solution at the same concentration as Aminoleban EN was defined as 3.5 (measured by a previously described gustatory sensation method). The bitterness threshold of a BCAA standard solution without added aroma was estimated to be 1.87, while those of BCAA solutions containing green-tea, coffee, apple, vanilla, or strawberry aromas were 2.02, 1.98, 2.35, 2.40 and 2.87, respectively, when evaluated by the probit method. This shows that the addition of an aroma can elevate the bitterness threshold in human volunteers. The green-tea and coffee aromas predominantly evoked bitterness, while the vanilla aroma predominantly evoked sweetness. Apple and strawberry aromas evoked both sweetness and sourness, with the apple aroma having stronger sourness and the strawberry aroma stronger sweetness. Thus, a 'sweet' aroma suppresses the bitterness of BCAA, with coexisting sourness also participating in the bitterness inhibition. [Abstract/Link to Full Text]

Bi Y, Mao S, Gan L, Li Y, Wang C, Xu N, Zheng Y, Cheng Q, Hou S
A controlled porosity osmotic pump system with biphasic release of theophylline.
Chem Pharm Bull (Tokyo). 2007 Nov;55(11):1574-80.
A controlled porosity osmotic pump system with biphasic release of theophylline was developed for the nocturnal therapy of asthma. The developed system was composed of a tablet-in-tablet (TNT) core and a controlled porosity coating membrane. Release pattern of the developed system was influenced by amount of pore former (18.2-45.5%, w/w of polymer), weight gain (16-26 mg per tablet) of the coating membrane and osmotic agents used in inner layer of the TNT core. When sodium phosphate and sodium chloride were selected as the osmotic agents in inner and outer layer of the TNT core respectively, target release profile was obtained with coating solution cellulose acetate-polyethylene glycol 400-diethyl phthalate (54.5-36.4-9.1%, w/w) at a weight gain of 16-22 mg per tablet. To examine the mechanism of drug release, release profiles of osmotic agents, micro-environmental osmotic pressure and micro-environmental pH of the formulation during dissolution were studied. Micro-environmental osmotic pressure decreased and micro-environmental pH increased continuously during the whole dissolution process, theophylline release was dominated by the successive dissolution of sodium chloride and sodium phosphate. Theophylline solubility increased as environmental pH exceeded 10.8. At the last stage of the biphasic release, micro-environmental pH in the developed formulation reached 10.9, and theophylline release was promoted by its elevated solubility despite of the decrease of micro-environmental osmotic pressure in the developed formulaiton. [Abstract/Link to Full Text]

Fukui A, Fujii R, Yonezawa Y, Sunada H
Analysis of the release process of phenylpropanolamine hydrochloride from ethylcellulose matrix granules IV.(1)) Evaluation of the controlled release properties for in vivo and in vitro release systems.
Chem Pharm Bull (Tokyo). 2007 Nov;55(11):1569-73.
In the pharmaceutical preparation of a controlled release drug, it is very important and necessary to understand the release properties. The dissolution test is a very important and useful method for understanding and predicting drug-release properties. It was readily confirmed in the previous paper that the release process could be assessed quantitatively by a combination of the square-root time law and cube-root law equations for ethylcellulose (EC) matrix granules of phenylpropanolamine hydrochloride (PPA). In this paper EC layered granules were used in addition to EC matrix. The relationship between release property and the concentration of PPA in plasma after administration using beagle dogs were examined. Then it was confirmed that the correlativity for EC layered granules and EC matrix were similar each other. Therefore, it was considered that the dissolution test is useful for prediction of changes in concentration of PPA in the blood with time. And it was suggested that EC layered granules were suitable as a controlled release system as well as EC matrix. [Abstract/Link to Full Text]

Yue PF, Yuan HL, Li XY, Yang M, Zhu WF, Xiao XH
Development and optimization of intravenous puerarin emulsions formation by a novel complex-phase inversion-homogenization technology.
Chem Pharm Bull (Tokyo). 2007 Nov;55(11):1563-8.
To decrease the hemolysis side effect of puerarin, a novel approach of Complex-Phase Inversion-Homogenization (CPIH) Technology was established to prepare intravenous puerarin emulsions. Preparation of puerarin submicron emulsion were optimized by central composite design, and the physicochemical properties were evaluated. Puerarin phospholipid complexs prepared by puerarin and phospholipids at a ratio of 1 : 1.2. In order to improve the product quality, a central composite design was applied to optimize the critical process variables, such as emulsification time, stirring velocity and homogenization press, and the results were modeled statistically. Puerarin phospholipid complexs prepared were identified by fourier transform infrared spectrophotometry. The datas showed that the parameters had great effect on the response values. The particle size, span of dispersity and entrapment efficiency of puerarin emulsions prepared using the optimal parameters settings were 218.23 nm, 0.6284 and 87.32%, respectively. These meant that over 87% of the drug was located at the surfactant interface and oil droplet, the concentration of puerarin in aqueous was rarely. And the puerarin emulsion prepared by CPIH technology were sufficient stable for 90 d. The CPIH technology can be used as a general formulation principle for drugs which are slightly soluble in water and poorly soluble in oils. [Abstract/Link to Full Text]

Oshima T, Sonoda R, Ohkuma M, Sunada H
Preparation of rapidly disintegrating tablets containing itraconazole solid dispersions.
Chem Pharm Bull (Tokyo). 2007 Nov;55(11):1557-62.
The disintegratability of tablets prepared from two types of solid dispersions containing the water-soluble polymer TC-5 and the enteric polymer HP-55 as an excipient were compared. The disintegratability was better in the tablets of solid dispersions containing non-water-soluble HP-55 than those containing TC-5. In consideration of the dissolubility of solid dispersions containing HP-55, the mean diameter of the solid dispersion (coating powder) must be controlled to 120 microm or less, but as this markedly increases the adhesion/aggregation tendency of the particles (angle of repose: 47 degrees ), control of the adhesion/aggregation tendency emerged as another problem. Therefore, surface-modification was performed in a high-speed agitating granulator using 0.1% light anhydrous silicic acid as a surface modifier, and marked improvement in the flowability was observed. This made continuous tableting using a rotary tablet machine possible even with the poorly flowable solid dispersions. Also, in tableting of the solid dispersions, no recrystallization of amorphous itraconazole by the tableting pressure was observed, and the tablets maintained satisfactory dissolubility. Moreover, it was possible to obtain the rapidly disintegrating tablets with very satisfactory properties, i.e., a tablet hardness of 30 N or higher and a disintegration time of 30 s or less, by the addition of croscarmellose as a disintegrant at 2% to the surface-modified solid dispersion and selection of the tableting pressure at 4.5 kN. [Abstract/Link to Full Text]

Askal H, Refaat I, Darwish I, Marzouq M
Evaluation of N-bromosuccinimide as a new analytical reagent for the spectrophotometric determination of fluoroquinolone antibiotics.
Chem Pharm Bull (Tokyo). 2007 Nov;55(11):1551-6.
Analytical studies were carried out, for the first time, to evaluate the use of N-bromosuccinimide (NBS) as an analytical reagent for the spectrophotometric determination of eleven therapeutically important fluoroquinolone antibiotics (FQA). The procedures involved the reaction of the FQA with NBS and subsequent measurement of the excess NBS by its reaction with p-phenylenediamine (PDA) to give a violet colored product that was measured at 530 nm. Different variables affecting the reaction (concentration of NBS, concentration of PDA, pH of reaction medium, reaction time, and the diluting solvents) were carefully studied and optimized. The molar ratio and mechanism of the reaction between each of the studied FQA with NBS were proposed using UV-vis, IR, and NMR techniques. Under the optimum reaction conditions, the analytical method was developed and validated. Beer's law was obeyed in the general concentration range of 3-25 microg/ml. The assay limits of detection and quantitation were 0.33-1.29 and 1.10-4.31 microg/ml, respectively. The precision of the method was satisfactory; the values of relative standard deviations did not exceed 2%. The proposed method was successfully applied to the analysis of the investigated FQA in their pure and pharmaceutical dosage forms without interference from the common excipients (label claim values were 99.85-100.17+/-0.13-0.59%). Interference from ascorbic acid, that is co-formulated as a stabilizer for the ampoule form, was avoided by its pre-oxidation with potassium bromate before applying the analytical procedure. The results obtained by the proposed method were comparable with those obtained by the official and reported methods. [Abstract/Link to Full Text]

Xu L, Li SM, Sunada H
Preparation and evaluation of Ibuprofen solid dispersion systems with kollidon particles using a pulse combustion dryer system.
Chem Pharm Bull (Tokyo). 2007 Nov;55(11):1545-50.
Solid dispersions (SDs) of ibuprofen (IBU) were prepared with four carriers: Kollidon 25, Kollidon 30, Kollidon VA64, and Kollidon CL, using a newly developed pulse combustion dryer system, HYPULCON. Physicochemical properties of the SDs obtained were investigated by differential scanning calorimetry (DSC), powder X-ray diffraction (PXRD), scanning electron microscope (SEM), and Fourier transformation IR spectroscopy (FT-IR). Powder X-ray diffraction (PXRD) showed that the crystal diffraction peaks of IBU in SDs disappeared completely, and in differential scanning calorimetry (DSC) curves, the endothermic peaks of IBU in SDs were not observed. Fourier transformation IR spectroscopy (FT-IR) proved that interactions between the drug and carrier existed. These findings demonstrated that IBU changed to an amorphous form in the SDs with the four carriers using the pulse combustion dryer system. The dissolution property of IBU in the SDs was markedly enhanced. The dissolution test showed that after 5 min of dissolution, the concentrations of IBU in the SDs with Kollidon CL as the carrier was 43.81 mug/ml, corresponding to 13.0 times that of pure IBU. So, it is demonstrated that the pulse combustion dryer system is very useful for preparing SDs of IBU with Kollidon of different grades as carriers. [Abstract/Link to Full Text]

Hashimoto M, Kato YH, Hatanaka Y
Selective hydrogenation of alkene in (3-trifluoromethyl) phenyldiazirine photophor with Wilkinson's catalyst for photoaffinity labeling.
Chem Pharm Bull (Tokyo). 2007 Oct;55(10):1540-3.
Selective hydrogenation of carbon-carbon double bond in the presence of nitrogen-nitrogen double bond in (3-trifluoromethyl) phenyldiazirine achieved with Wilkinson's catalyst. [Abstract/Link to Full Text]

Ito T, Abe N, Ali Z, Oyama M, Tanaka T, Murata J, Darnaedi D, Iinuma M
Resveratrol tetramers with a C6-C3 or a C1 unit from Upuna borneensis.
Chem Pharm Bull (Tokyo). 2007 Oct;55(10):1535-9.
Investigation of the chemical constituents in the stem of Upuna borneensis (Dipterocarpaceae) resulted in the isolation of three new resveratrol derivatives, upunaphenols L (1), M (2) (resveratrol tetramers with a C(6)-C(3) unit) and N (3) (resveratrol tetramer with a C(1) unit). The structures have the same partial structure as vaticanol B (4). Upunaphenols L and M are new complex polyphenol compounds, lignostilbene. Their structures were determined by spectroscopic analysis including two dimensional NMR. Upunaphenol M was found to be an artifact generated by silica gel catalyzed methanolysis of 1. [Abstract/Link to Full Text]

Guo Y, Li Y, Xu J, Li N, Yamakuni T, Ohizumi Y
Clerodane diterpenoids and flavonoids with NGF-potentiating activity from the aerial parts of Baccharis gaudichaudiana.
Chem Pharm Bull (Tokyo). 2007 Oct;55(10):1532-4.
A new clerodane diterpene, 15-hydroxy-16-acetoxy-ent-clerod-3-en-18-oic acid (1), together with three known clerodane diterpenes (2-4) and three known flavones (5-7), were isolated from the aerial parts of Baccharis gaudichaudiana. Their structures were elucidated on the basis of spectroscopic analysis. Compounds 2, 3, and 5 showed enhancing activity of nerve growth factor (NGF)-induced neurite outgrowth in PC 12D cells. [Abstract/Link to Full Text]

Kodai T, Umebayashi K, Nakatani T, Ishiyama K, Noda N
Compositions of royal jelly II. Organic acid glycosides and sterols of the royal jelly of honeybees (Apis mellifera).
Chem Pharm Bull (Tokyo). 2007 Oct;55(10):1528-31.
Two organic acid glycosides (1, 2) and 16 sterols were isolated from the royal jelly of honeybees (Apis mellifera). The former two were monoglucosides of 10-hydroxy-2E-decenoic and 10-hydroxydecanoic acids. They are the first examples of glycosides isolated from royal jelly. The latter 16 were sterols mainly composed of 28 or 29 carbons. Among them, four compounds were new isofucosterol derivatives, and their structures were characterized as (24Z)-stigmasta-5,24(28)-dien-3beta-ol-7-one (3), (24Z)-stigmasta-5,24(28)-diene-3beta,7beta-diol (4), (24Z)-stigmasta-5,24(28)-diene-3beta,7alpha-diol (5), and (24Z)-stigmast-24(28)-ene-3beta,5alpha,6beta-triol (6) on the basis of various NMR spectroscopic data. [Abstract/Link to Full Text]

Ko SR, Suzuki Y, Suzuki K, Choi KJ, Cho BG
Marked production of ginsenosides Rd, F2, Rg3, and compound K by enzymatic method.
Chem Pharm Bull (Tokyo). 2007 Oct;55(10):1522-7.
The hydrolysis of protopanaxadiol-type saponin mixture by various glycoside hydrolases was examined. Among these enzymes, crude preparations of lactase from Aspergillus oryzae, beta-galactosidase from A. oryzae, and cellulase from Trichoderma viride were found to produce ginsenoside F(2) [3-O-(beta-D-glucopyranosyl)-20-O-beta-D-glucopyranosyl-20(S)-protopanaxadiol], compound K [20-O-beta-D-glucopyranosyl-20(S)-protopanaxadiol], and ginsenoside Rd {3-O-[beta-D-glucopyranosyl-(1-->2)-beta-D-glucopyranosyl]-20-O-beta-D-glucopyranosyl-20(S)-protopanaxadiol}, respectively, from protopanaxadiol-type saponin mixture in large quantities. Moreover, the crude preparation of lactase from Penicillium sp. having a high producing activity of ginsenoside Rh(1) (6-O-beta-D-glucopyranosyl-20(S)-protopanaxatriol) from protopanaxatriol-type saponin mixture gave ginsenoside Rd as a main product, ginsenoside Rg(3) {3-O-[beta-D-glucopyranosyl-(1-->2)-beta-D-glucopyranosyl]-20(S)-protopanaxadiol}, and compound K from protopanaxadiol-type saponin mixture. The hydrolytic pathways of ginsenosides Rb(1), Rb(2), and Rc to ginsenosides Rd, Rg(3), and F(2), and compound K by crude preparations of four glycoside hydrolases were also studied. This is the first report on the enzymatic preparation of an intestinal bacterial metabolite, ginsenoside F(2), in quantity, and a considerable amount of a minor saponin, ginsenoside Rg(3), from a protopanaxadiol-type saponin mixture. [Abstract/Link to Full Text]

Ohyama K, Suzuki M, Masuda K, Yoshida S, Muranaka T
Chemical phenotypes of the hmg1 and hmg2 mutants of Arabidopsis demonstrate the in-planta role of HMG-CoA reductase in triterpene biosynthesis.
Chem Pharm Bull (Tokyo). 2007 Oct;55(10):1518-21.
Plants produce a wide variety of cyclic triterpenes, such as sterols and triterpenoids, which are the major products of the mevalonate (MVA) pathway. It is important to understand the physiological functions of HMG-CoA reductase (HMGR) because HMGR is the rate-limiting enzyme in the MVA pathway. We have previously isolated Arabidopsis mutants in HMG1 and HMG2. Although the biochemical function of HMGR2 has been thought to be almost equal to that of HMGR1, based on similarities in their sequences, the phenotypes of mutants in these genes are quite different. Whereas hmg2 shows no abnormal phenotype under normal growth conditions, hmg1 shows pleiotropic phenotypes, including dwarfing, early senescence, and male sterility. We previously postulated that the 50% decrease in the sterol content of hmg1, as compared to that in the wild type, was a cause of these phenotypes, but comprehensive triterpene profiles of these mutants had not yet been determined. Here, we present the triterpene profiles of hmg1 and hmg2. In contrast to hmg1, hmg2 showed a sterol content 15% lower than that of the wild type. A precise triterpenoid quantification using synthesized deuterated compounds of beta-amyrin (1), alpha-amyrin (2), and lupeol (3) showed that the levels of triterpenoids in hmg1 and hmg2 were 65% and 25% lower than in the wild type (WT), respectively. These results demonstrate that HMGR2 as well as HMGR1 is responsible for the biosynthesis of triterpenes in spite of the lack of visible phenotypes in hmg2. [Abstract/Link to Full Text]

Sekizaki H, Itoh K, Shibuya A, Toyota E, Tanizawa K
A facile synthesis of p- and m-(amidinomethyl)phenyl esters derived from amino acid and tryptic hydrolysis of these synthetic inverse substrates.
Chem Pharm Bull (Tokyo). 2007 Oct;55(10):1514-7.
A facile synthetic method for p- and m-(amidinomethyl)phenyl esters derived from a variety of amino acids is presented. We analyzed the kinetic behavior of trypsin towards these synthetic esters, which are inverse substrates. The substituent (meta- and para-isomers) and isosteric effects of (amidinomethyl)phenyl esters are discussed. [Abstract/Link to Full Text]

Herath W, Reddy N, Khan IA
Microbial metabolism. Part 8. The pyranocoumarin, decursin.
Chem Pharm Bull (Tokyo). 2007 Oct;55(10):1512-3.
Microbial transformation of the cancer chemopreventive agent, decursin (1) with Sepedonium chrysospermem (ATCC 13378) yielded two metabolites, (+)-decursinol (2) and (-)-cis-decursidinol (3). The structures were established by spectroscopic data. [Abstract/Link to Full Text]

Nakamura S, Li X, Matsuda H, Ninomiya K, Morikawa T, Yamaguti K, Yoshikawa M
Bioactive constituents from Chinese natural medicines. XXVI. Chemical structures and hepatoprotective effects of constituents from roots of Rhodiola sachalinensis.
Chem Pharm Bull (Tokyo). 2007 Oct;55(10):1505-11.
The methanolic extract from the roots of Rhodiola sachalinensis was found to show a protective effect on D-galactosamine-induced cytotoxicity in primary cultured mouse hepatocytes. From the methanolic extract, five new glycosides, two monoterpene glycosides, two flavonol bisdesmosides, and a cyanogenic glycoside, were isolated together with 34 known compounds. The structures of new constituents were elucidated on the basis of chemical and physicochemical evidence. In addition, the principal constituents, sachalosides III and IV, rhodiosin, and trans-caffeic acid, displayed hepatoprotective effects. [Abstract/Link to Full Text]

Noguchi T, Tanaka N, Nishimata T, Goto R, Hayakawa M, Sugidachi A, Ogawa T, Asai F, Fujimoto K
Cinnamylindoline derivatives: synthesis and factor Xa (FXa) inhibitory activities.
Chem Pharm Bull (Tokyo). 2007 Oct;55(10):1494-504.
A series of cinnamylindoline derivatives were synthesized, and their factor Xa (FXa) inhibitory activities and selectivity over trypsin were evaluated. Among them, some novel derivatives showed potent FXa inhibitory activities and good selectivity over trypsin. Especially, (E)-2-{5-[1-(acetimidoyl)piperidin-4-yloxy]-2-[2-(5-amidino-2-hydroxyphenyl)ethen-1-yl]indolin-1-ylsulfonyl}acetic acid (22f) having 2-hydroxycinnamyl moiety exhibited the most potent FXa inhibitory activity in vitro. Furthermore, 22f also exhibited potent anticoagulant activities in vitro. [Abstract/Link to Full Text]

Osa Y, Ida Y, Fujii H, Nemoto T, Hasebe K, Momen S, Mochizuki H, Nagase H
Catalytic aerobic oxidation of nor-binaltorphimine (nor-BNI) analogs without 4,5-epoxy bridge affords a more selective ligand for kappa opioid receptor than the representative kappa antagonist nor-BNI.
Chem Pharm Bull (Tokyo). 2007 Oct;55(10):1489-93.
An analog of nor-binaltorphimine (nor-BNI) without the 4,5-epoxy bridge, 17,17'-bis(cyclopropylmethyl)-6,6',7,7'-tetrahydro-6,6'-imino-14beta,14'alpha-dihydroxy-3,3'-dimethoxy-7,7'-bimorphinan (4), which was the precursor of the designed compound 1 as a selective kappa(3) opioid receptor antagonist, was catalytically oxidized with oxygen in the presence of platinum to give the 5'-oxo derivative 3 with some other oxidized products. Morphinan derivatives without the 4,5-epoxy moiety were labile to oxygen, although the corresponding 4,5-epoxymorphinan derivatives resisted aerobic oxidation. One of the oxidized nor-BNI analogs without 4,5-epoxy bridge, compound 18, showed high affinity and selectivity for kappa opioid receptor. [Abstract/Link to Full Text]

Ghoneim EM
Electroreduction of the muscle relaxant drug dantrolene sodium at the mercury electrode and its determination in bulk form and pharmaceutical formulation.
Chem Pharm Bull (Tokyo). 2007 Oct;55(10):1483-8.
The electroreduction of the muscle relaxant drug dantrolene sodium at the mercury electrode has been studied in the Britton-Robinson universal buffer of pH 2.5-11.5 containing 20% (v/v) methanol by means of dc-polarography, cyclic voltammetry and controlled-potential coulometry. Its reduction took place via three irreversible cathodic steps in solutions of pH < or =6, two steps in solutions of 6<pH<10 and a single step at pH values > or =10 through the consumption of 10, 8 or 4 electrons, respectively. This behavior was attributed to the reduction of NO(2) group (1st and 2nd steps at pH < or =6 or the single step at pH > or =10) and the -CH=N- double bond (3rd step at pH <10). Two polarographic procedures (direct current and differential-pulse modes) and three adsorptive cathodic stripping voltammetric procedures (linear-sweep, differential-pulse and square-wave modes) were described and successfully applied for quantification of dantrolene sodium in its bulk form and in pharmaceutical formulation (Dantrolex tablets). [Abstract/Link to Full Text]

El-Halawany AM, Chung MH, Nakamura N, Ma CM, Nishihara T, Hattori M
Estrogenic and anti-estrogenic activities of Cassia tora phenolic constituents.
Chem Pharm Bull (Tokyo). 2007 Oct;55(10):1476-82.
Through an estrogenic activity bioassay-guided fractionation of the 70% ethanolic extract of Cassia tora seeds two new phenolic triglucosides, torachrysone 8-O-[beta-D-glucopyranosyl(1-->3)-O-beta-D-glucopyranosyl(1-->6)-O-beta-D-glucopyranoside] (1) and toralactone 9-O-[beta-D-glucopyranosyl-(1-->3)-O-beta-D-glucopyranosyl-(1-->6)-O-beta-D-glucopyranoside] (2), along with seven known compounds were isolated. The structures of the new compounds were elucidated on the basis of spectroscopic and chemical evidence. The estrogenic activity of the fractions and the isolated compounds were investigated using the estrogen-dependent proliferation of MCF-7 cells. In addition, the yeast two hybrid assay expressing estrogen receptor alpha (ERalpha) and beta (ERbeta) and the ERalpha competitor screening assay (ligand binding screen) were used to verify the binding affinities of the isolated compounds to ER. Furthermore, a naringinase pre-treatment of the 70% alcoholic extract of Cassia tora seeds resulted in a significant increase in its estrogenic activity. From the naringinase pre-treated extract six compounds were isolated, among which 6-hydroxymusizin and aurantio-obtusin showed the most potent estrogenic activity, while torachrysone, rubrofusarin and toralactone showed a significant anti-estrogenic activity. Finally, the structure requirements responsible for the estrogenic activity of the isolated compounds were studied by investigating the activity of several synthetic compounds and chemically modifying the isolated compounds. The basic nucleus 1,3,8-trihyroxynaphthalene (T(3)HN) was found to play a principal role in the binding affinity of these compounds to ER. [Abstract/Link to Full Text]

Chen SP, Su JH, Ahmed AF, Dai CF, Wu YC, Sheu JH
Xeniaphyllane-derived terpenoids from the formosan soft coral Sinularia gibberosa.
Chem Pharm Bull (Tokyo). 2007 Oct;55(10):1471-5.
New xeniaphyllane-derived metabolites (1-7) were isolated from the EtOAc extract of the Formosan soft coral Sinularia gibberosa. The structures and relative configurations of these compounds were elucidated on the basis of extensive spectroscopic analysis (including 2D NMR) and by comparison of their spectral data with those of related compounds. In vitro cytotoxic evaluation of the above metabolites towards a limited panel of cancer cell lines is also described. [Abstract/Link to Full Text]

Recent Articles in The Journal of Experimental Medicine

Schroen B, Leenders JJ, van Erk A, Bertrand AT, van Loon M, van Leeuwen RE, Kubben N, Duisters RF, Schellings MW, Janssen BJ, Debets JJ, Schwake M, Hřydal MA, Heymans S, Saftig P, Pinto YM
Lysosomal integral membrane protein 2 is a novel component of the cardiac intercalated disc and vital for load-induced cardiac myocyte hypertrophy.
J Exp Med. 2007 May 14;204(5):1227-35.
The intercalated disc (ID) of cardiac myocytes is emerging as a crucial structure in the heart. Loss of ID proteins like N-cadherin causes lethal cardiac abnormalities, and mutations in ID proteins cause human cardiomyopathy. A comprehensive screen for novel mechanisms in failing hearts demonstrated that expression of the lysosomal integral membrane protein 2 (LIMP-2) is increased in cardiac hypertrophy and heart failure in both rat and human myocardium. Complete loss of LIMP-2 in genetically engineered mice did not affect cardiac development; however, these LIMP-2 null mice failed to mount a hypertrophic response to increased blood pressure but developed cardiomyopathy. Disturbed cadherin localization in these hearts suggested that LIMP-2 has important functions outside lysosomes. Indeed, we also find LIMP-2 in the ID, where it associates with cadherin. RNAi-mediated knockdown of LIMP-2 decreases the binding of phosphorylated beta-catenin to cadherin, whereas overexpression of LIMP-2 has the opposite effect. Collectively, our data show that LIMP-2 is crucial to mount the adaptive hypertrophic response to cardiac loading. We demonstrate a novel role for LIMP-2 as an important mediator of the ID. [Abstract/Link to Full Text]

Péron S, Pan-Hammarström Q, Imai K, Du L, Taubenheim N, Sanal O, Marodi L, Bergelin-Besançon A, Benkerrou M, de Villartay JP, Fischer A, Revy P, Durandy A
A primary immunodeficiency characterized by defective immunoglobulin class switch recombination and impaired DNA repair.
J Exp Med. 2007 May 14;204(5):1207-16.
Immunoglobulin class switch recombination (CSR) deficiencies are rare primary immunodeficiencies, characterized by a lack of switched isotype (IgG, IgA, or IgE) production, variably associated with abnormal somatic hypermutation (SHM). Deficiencies in CD40 ligand, CD40, activation-induced cytidine deaminase, and uracil-N-glycosylase may account for this syndrome. We previously described another Ig CSR deficiency condition, characterized by a defect in CSR downstream of the generation of double-stranded DNA breaks in switch (S) mu regions. Further analysis performed with the cells of five affected patients showed that the Ig CSR deficiency was associated with an abnormal formation of the S junctions characterized by microhomology and with increased cell radiosensitivity. In addition, SHM was skewed toward transitions at G/C residues. Overall, these findings suggest that a unique Ig CSR deficiency phenotype could be related to an as-yet-uncharacterized defect in a DNA repair pathway involved in both CSR and SHM events. [Abstract/Link to Full Text]

Arnold L, Henry A, Poron F, Baba-Amer Y, van Rooijen N, Plonquet A, Gherardi RK, Chazaud B
Inflammatory monocytes recruited after skeletal muscle injury switch into antiinflammatory macrophages to support myogenesis.
J Exp Med. 2007 May 14;204(5):1057-69.
Macrophages (MPs) are important for skeletal muscle regeneration in vivo and may exert beneficial effects on myogenic cell growth through mitogenic and antiapoptotic activities in vitro. However, MPs are highly versatile and may exert various, and even opposite, functions depending on their activation state. We studied monocyte (MO)/MP phenotypes and functions during skeletal muscle repair. Selective labeling of circulating MOs by latex beads in CX3CR1(GFP/+) mice showed that injured muscle recruited only CX3CR1(lo)/Ly-6C(+) MOs from blood that exhibited a nondividing, F4/80(lo), proinflammatory profile. Then, within muscle, these cells switched their phenotype to become proliferating antiinflammatory CX3CR1(hi)/Ly-6C(-) cells that further differentiated into F4/80(hi) MPs. In vitro, phagocytosis of muscle cell debris induced a switch of proinflammatory MPs toward an antiinflammatory phenotype releasing transforming growth factor beta1. In co-cultures, inflammatory MPs stimulated myogenic cell proliferation, whereas antiinflammatory MPs exhibited differentiating activity, assessed by both myogenin expression and fusion into myotubes. Finally, depletion of circulating MOs in CD11b-diphtheria toxin receptor mice at the time of injury totally prevented muscle regeneration, whereas depletion of intramuscular F4/80(hi) MPs at later stages reduced the diameter of regenerating fibers. In conclusion, injured skeletal muscle recruits MOs exhibiting inflammatory profiles that operate phagocytosis and rapidly convert to antiinflammatory MPs that stimulate myogenesis and fiber growth. [Abstract/Link to Full Text]

Feldhahn N, Henke N, Melchior K, Duy C, Soh BN, Klein F, von Levetzow G, Giebel B, Li A, Hofmann WK, Jumaa H, Müschen M
Activation-induced cytidine deaminase acts as a mutator in BCR-ABL1-transformed acute lymphoblastic leukemia cells.
J Exp Med. 2007 May 14;204(5):1157-66.
The Philadelphia chromosome (Ph) encoding the oncogenic BCR-ABL1 kinase defines a subset of acute lymphoblastic leukemia (ALL) with a particularly unfavorable prognosis. ALL cells are derived from B cell precursors in most cases and typically carry rearranged immunoglobulin heavy chain (IGH) variable (V) region genes devoid of somatic mutations. Somatic hypermutation is restricted to mature germinal center B cells and depends on activation-induced cytidine deaminase (AID). Studying AID expression in 108 cases of ALL, we detected AID mRNA in 24 of 28 Ph(+) ALLs as compared with 6 of 80 Ph(-) ALLs. Forced expression of BCR-ABL1 in Ph(-) ALL cells and inhibition of the BCR-ABL1 kinase showed that aberrant expression of AID depends on BCR-ABL1 kinase activity. Consistent with aberrant AID expression in Ph(+) ALL, IGH V region genes and BCL6 were mutated in many Ph(+) but unmutated in most Ph(-) cases. In addition, AID introduced DNA single-strand breaks within the tumor suppressor gene CDKN2B in Ph(+) ALL cells, which was sensitive to BCR-ABL1 kinase inhibition and silencing of AID expression by RNA interference. These findings identify AID as a BCR-ABL1-induced mutator in Ph(+) ALL cells, which may be relevant with respect to the particularly unfavorable prognosis of this leukemia subset. [Abstract/Link to Full Text]

Humphreys IR, de Trez C, Kinkade A, Benedict CA, Croft M, Ware CF
Cytomegalovirus exploits IL-10-mediated immune regulation in the salivary glands.
J Exp Med. 2007 May 14;204(5):1217-25.
The salivary glands represent a major site of cytomegalovirus replication and transmission to other hosts. Despite control of viral infection by strong T cell responses in visceral organs cytomegalovirus replication continues in the salivary glands of mice, suggesting that the virus exploits the mucosal microenvironment. Here, we show that T cell immunity in the salivary glands is limited by the induction of CD4 T cells expressing the regulatory cytokine interleukin (IL)-10. Blockade of IL-10 receptor (IL-10R) with an antagonist antibody dramatically reduced viral load in the salivary glands, but not in the spleen. The mucosa-specific protection afforded by IL-10R blockade was associated with an increased accumulation of CD4 T cells expressing interferon gamma, suggesting that IL-10R signaling limits effector T cell differentiation. Consistent with this, an agonist antibody targeting the tumor necrosis factor receptor superfamily member OX40 (TNFRSF4) enhanced effector T cell differentiation and increased the number of interferon gamma-producing T cells, thus limiting virus replication in the salivary glands. Collectively, the results indicate that modulating effector T cell differentiation can counteract pathogen exploitation of the mucosa, thus limiting persistent virus replication and transmission. [Abstract/Link to Full Text]

Peixoto A, Evaristo C, Munitic I, Monteiro M, Charbit A, Rocha B, Veiga-Fernandes H
CD8 single-cell gene coexpression reveals three different effector types present at distinct phases of the immune response.
J Exp Med. 2007 May 14;204(5):1193-205.
To study in vivo CD8 T cell differentiation, we quantified the coexpression of multiple genes in single cells throughout immune responses. After in vitro activation, CD8 T cells rapidly express effector molecules and cease their expression when the antigen is removed. Gene behavior after in vivo activation, in contrast, was quite heterogeneous. Different mRNAs were induced at very different time points of the response, were transcribed during different time periods, and could decline or persist independently of the antigen load. Consequently, distinct gene coexpression patterns/different cell types were generated at the various phases of the immune responses. During primary stimulation, inflammatory molecules were induced and down-regulated shortly after activation, generating early cells that only mediated inflammation. Cytotoxic T cells were generated at the peak of the primary response, when individual cells simultaneously expressed multiple killer molecules, whereas memory cells lost killer capacity because they no longer coexpressed killer genes. Surprisingly, during secondary responses gene transcription became permanent. Secondary cells recovered after antigen elimination were more efficient killers than cytotoxic T cells present at the peak of the primary response. Thus, primary responses produced two transient effector types. However, after boosting, CD8 T cells differentiate into long-lived killer cells that persist in vivo in the absence of antigen. [Abstract/Link to Full Text]

McCarthy C, Shepherd D, Fleire S, Stronge VS, Koch M, Illarionov PA, Bossi G, Salio M, Denkberg G, Reddington F, Tarlton A, Reddy BG, Schmidt RR, Reiter Y, Griffiths GM, van der Merwe PA, Besra GS, Jones EY, Batista FD, Cerundolo V
The length of lipids bound to human CD1d molecules modulates the affinity of NKT cell TCR and the threshold of NKT cell activation.
J Exp Med. 2007 May 14;204(5):1131-44.
CD1d-restricted lymphocytes recognize a broad lipid range. However, how CD1d-restricted lymphocytes translate T cell receptor (TCR) recognition of lipids with similar group heads into distinct biological responses remains unclear. Using a soluble invariant NKT (iNKT) TCR and a newly engineered antibody specific for alpha-galactosylceramide (alpha-GalCer)-human CD1d (hCD1d) complexes, we measured the affinity of binding of iNKT TCR to hCD1d molecules loaded with a panel of alpha-GalCer analogues and assessed the rate of dissociation of alpha-GalCer and alpha-GalCer analogues from hCD1d molecules. We extended this analysis by studying iNKT cell synapse formation and iNKT cell activation by the same panel of alpha-GalCer analogues. Our results indicate the unique role of the lipid chain occupying the hCD1d F' channel in modulating TCR binding affinity to hCD1d-lipid complexes, the formation of stable immunological synapse, and cell activation. These data are consistent with previously described conformational changes between empty and loaded hCD1d molecules (Koch, M., V.S. Stronge, D. Shepherd, S.D. Gadola, B. Mathew, G. Ritter, A.R. Fersht, G.S. Besra, R.R. Schmidt, E.Y. Jones, and V. Cerundolo. 2005. Nat. Immunol 6:819-826), suggesting that incomplete occupation of the hCD1d F' channel results in conformational differences at the TCR recognition surface. This indirect effect provides a general mechanism by which lipid-specific lymphocytes are capable of recognizing both the group head and the length of lipid antigens, ensuring greater specificity of antigen recognition. [Abstract/Link to Full Text]

Asperti-Boursin F, Real E, Bismuth G, Trautmann A, Donnadieu E
CCR7 ligands control basal T cell motility within lymph node slices in a phosphoinositide 3-kinase-independent manner.
J Exp Med. 2007 May 14;204(5):1167-79.
The molecular mechanisms responsible for the sustained basal motility of T cells within lymph nodes (LNs) remain elusive. To study T cell motility in a LN environment, we have developed a new experimental system based on slices of LNs that allows the assessment of T cell trafficking after adoptive transfer or direct addition of T cells to the slice. Using this experimental system, we show that T cell motility is highly sensitive to pertussis toxin and strongly depends on CCR7 and its ligands. Our results also demonstrate that, despite its established role in myeloid cell locomotion, phosphoinositide 3-kinase (PI3K) activity does not contribute to the exploratory behavior of the T lymphocytes within LN slices. Likewise, although PI3K activation is detectable in chemokine-treated T cells, PI3K plays only a minor role in T cell polarization and migration in vitro. Collectively, our results suggest that the common amplification system that, in other cells, facilitates large phosphatidylinositol 3,4,5-trisphosphate increases at the plasma membrane is absent in T cells. We conclude that T cell motility within LNs is not an intrinsic property of T lymphocytes but is driven in a PI3K-independent manner by the lymphoid chemokine-rich environment. [Abstract/Link to Full Text]

Zhang X, Deriaud E, Jiao X, Braun D, Leclerc C, Lo-Man R
Type I interferons protect neonates from acute inflammation through interleukin 10-producing B cells.
J Exp Med. 2007 May 14;204(5):1107-18.
Newborns and infants are highly susceptible to viral and bacterial infections, but the underlying mechanism remains poorly understood. We show that neonatal B cells effectively control the production of proinflammatory cytokines by both neonatal plasmacytoid and conventional dendritic cells, in an interleukin (IL) 10-dependent manner, after Toll-like receptor (TLR) 9 triggering. This antiinflammatory property of neonatal B cells may extend to other TLR agonists (Pam3CSK4, lipopolysaccharide, and R848) and viruses. In the absence of B cells or of CD5(+) B cell subsets, neonatal mice developed stronger inflammatory responses and became lethally susceptible to CpG challenge after galactosamine sensitization, whereas wild-type (WT) mice were resistant. Paradoxically, interferon (IFN)-alpha/beta enhanced the inflammatory response to CpG challenge in adult mice, whereas they helped to control neonatal acute inflammation by stimulating the secretion of IL-10 by neonatal B cells. Finally, WT neonatal B cells rescued IL-10(-/-) neonates from a lethal CpG challenge, whereas IFN-alpha/beta receptor-deficient B cells did not. Our results show that type I IFNs support a negative regulatory role of neonatal B cells on TLR-mediated inflammation, with important implications for neonatal inflammation and infection. [Abstract/Link to Full Text]

Kawagoe T, Sato S, Jung A, Yamamoto M, Matsui K, Kato H, Uematsu S, Takeuchi O, Akira S
Essential role of IRAK-4 protein and its kinase activity in Toll-like receptor-mediated immune responses but not in TCR signaling.
J Exp Med. 2007 May 14;204(5):1013-24.
Interleukin-1 receptor-associated kinase 4 (IRAK-4) was reported to be essential for the Toll-like receptor (TLR)- and T cell receptor (TCR)-mediated signaling leading to the activation of nuclear factor kappaB (NF-kappaB). However, the importance of kinase activity of IRAK family members is unclear. In this study, we investigated the functional role of IRAK-4 activity in vivo by generating mice carrying a knockin mutation (KK213AA) that abrogates its kinase activity. IRAK-4(KN/KN) mice were highly resistant to TLR-induced shock response. The cytokine production in response to TLR ligands was severely impaired in IRAK-4(KN/KN) as well as IRAK-4(-/-) macrophages. The IRAK-4 activity was essential for the activation of signaling pathways leading to mitogen-activated protein kinases. TLR-induced IRAK-4/IRAK-1-dependent and -independent pathways were involved in early induction of NF-kappaB-regulated genes in response to TLR ligands such as tumor necrosis factor alpha and IkappaBzeta. In contrast to a previous paper (Suzuki, N., S. Suzuki, D.G. Millar, M. Unno, H. Hara, T. Calzascia, S. Yamasaki, T. Yokosuka, N.J. Chen, A.R. Elford, et al. 2006. Science. 311:1927-1932), the TCR signaling was not impaired in IRAK-4(-/-) and IRAK-4(KN/KN) mice. Thus, the kinase activity of IRAK-4 is essential for the regulation of TLR-mediated innate immune responses. [Abstract/Link to Full Text]

Kang HR, Lee CG, Homer RJ, Elias JA
Semaphorin 7A plays a critical role in TGF-beta1-induced pulmonary fibrosis.
J Exp Med. 2007 May 14;204(5):1083-93.
Semaphorin (SEMA) 7A regulates neuronal and immune function. In these studies, we tested the hypothesis that SEMA 7A is also a critical regulator of tissue remodeling. These studies demonstrate that SEMA 7A and its receptors, plexin C1 and beta1 integrins, are stimulated by transforming growth factor (TGF)-beta(1) in the murine lung. They also demonstrate that SEMA 7A plays a critical role in TGF-beta(1)-induced fibrosis, myofibroblast hyperplasia, alveolar remodeling, and apoptosis. TGF-beta(1) stimulated SEMA 7A via a largely Smad 3-independent mechanism and stimulated SEMA 7A receptors, matrix proteins, CCN proteins, fibroblast growth factor 2, interleukin 13 receptor components, proteases, antiprotease, and apoptosis regulators via Smad 2/3-independent and SEMA 7A-dependent mechanisms. SEMA 7A also played an important role in the pathogenesis of bleomycin-induced pulmonary fibrosis. TGF-beta(1) and bleomycin also activated phosphatidylinositol 3-kinase (PI3K) and protein kinase B (PKB)/AKT via SEMA 7A-dependent mechanisms, and PKB/AKT inhibition diminished TGF-beta(1)-induced fibrosis. These observations demonstrate that SEMA 7A and its receptors are induced by TGF-beta(1) and that SEMA 7A plays a central role in a PI3K/PKB/AKT-dependent pathway that contributes to TGF-beta(1)-induced fibrosis and remodeling. They also demonstrate that the effects of SEMA 7A are not specific for transgenic TGF-beta(1), highlighting the importance of these findings for other fibrotic stimuli. [Abstract/Link to Full Text]

Zangani MM, Frřyland M, Qiu GY, Meza-Zepeda LA, Kutok JL, Thompson KM, Munthe LA, Bogen B
Lymphomas can develop from B cells chronically helped by idiotype-specific T cells.
J Exp Med. 2007 May 14;204(5):1181-91.
B cell lymphomas have been associated with chronic infections and autoimmunity. However, most lymphomas develop in the absence of any known chronic antigenic stimulation. B cells process their highly diversified endogenous immunoglobulin and present clonally unique variable-region idiotypic (Id) peptides on their major histocompatibility complex (MHC) class II molecules to Id-specific T cells. We show that B cells chronically helped by Id-specific Th2 cells developed into large B cell lymphomas with cytogenetic DNA aberrations. The lymphomas expressed high amounts of Id, MHC class II, CD80/86, and CD40 and bidirectionally collaborated with Th2 cells. Thus, MHC class II-presented Id peptides may represent a chronic self-antigenic stimulus for T cell-dependent lymphomagenesis. Eventually, B lymphomas grew independent of T cells. Thus, T cells do not only eliminate cancers as currently believed. In fact, Id-specific Th2 cells can induce B lymphomas. [Abstract/Link to Full Text]

Kim TW, Staschke K, Bulek K, Yao J, Peters K, Oh KH, Vandenburg Y, Xiao H, Qian W, Hamilton T, Min B, Sen G, Gilmour R, Li X
A critical role for IRAK4 kinase activity in Toll-like receptor-mediated innate immunity.
J Exp Med. 2007 May 14;204(5):1025-36.
IRAK4 is a member of IL-1 receptor (IL-1R)-associated kinase (IRAK) family and has been shown to play an essential role in Toll-like receptor (TLR)-mediated signaling. We recently generated IRAK4 kinase-inactive knock-in mice to examine the role of kinase activity of IRAK4 in TLR-mediated signaling pathways. The IRAK4 kinase-inactive knock-in mice were completely resistant to lipopolysaccharide (LPS)- and CpG-induced shock, due to impaired TLR-mediated induction of proinflammatory cytokines and chemokines. Although inactivation of IRAK4 kinase activity did not affect the levels of TLR/IL-1R-mediated nuclear factor kappaB activation, a reduction of LPS-, R848-, and IL-1-mediated mRNA stability contributed to the reduced cytokine and chemokine production in bone marrow-derived macrophages from IRAK4 kinase-inactive knock-in mice. Both TLR7- and TLR9-mediated type I interferon production was abolished in plasmacytoid dendritic cells isolated from IRAK4 knock-in mice. In addition, influenza virus-induced production of interferons in plasmacytoid DCs was also dependent on IRAK4 kinase activity. Collectively, our results indicate that IRAK4 kinase activity plays a critical role in TLR-dependent immune responses. [Abstract/Link to Full Text]

Michel ML, Keller AC, Paget C, Fujio M, Trottein F, Savage PB, Wong CH, Schneider E, Dy M, Leite-de-Moraes MC
Identification of an IL-17-producing NK1.1(neg) iNKT cell population involved in airway neutrophilia.
J Exp Med. 2007 May 14;204(5):995-1001.
Invariant natural killer T (iNKT) cells are an important source of both T helper type 1 (Th1) and Th2 cytokines, through which they can exert beneficial, as well as deleterious, effects in a variety of inflammatory diseases. This functional heterogeneity raises the question of how far phenotypically distinct subpopulations are responsible for such contrasting activities. In this study, we identify a particular set of iNKT cells that lack the NK1.1 marker (NK1.1(neg)) and secrete high amounts of interleukin (IL)-17 and low levels of interferon (IFN)-gamma and IL-4. NK1.1(neg) iNKT cells produce IL-17 upon synthetic (alpha-galactosylceramide [alpha-GalCer] or PBS-57), as well as natural (lipopolysaccharides or glycolipids derived from Sphingomonas wittichii and Borrelia burgdorferi), ligand stimulation. NK1.1(neg) iNKT cells are more frequent in the lung, which is consistent with a role in the natural immunity to inhaled antigens. Indeed, airway neutrophilia induced by alpha-GalCer or lipopolysaccharide instillation was significantly reduced in iNKT-cell-deficient Jalpha18(-/-) mice, which produced significantly less IL-17 in their bronchoalveolar lavage fluid than wild-type controls. Furthermore, airway neutrophilia was abolished by a single treatment with neutralizing monoclonal antibody against IL-17 before alpha-GalCer administration. Collectively, our findings reveal that NK1.1(neg) iNKT lymphocytes represent a new population of IL-17-producing cells that can contribute to neutrophil recruitment through preferential IL-17 secretion. [Abstract/Link to Full Text]

Shi L, Godfrey WR, Lin J, Zhao G, Kao PN
NF90 regulates inducible IL-2 gene expression in T cells.
J Exp Med. 2007 May 14;204(5):971-7.
Activation of T cells induces the production of T cell growth and survival factor interleukin (IL) 2. Regulatory T cells intrinsically fail to induce IL-2 expression upon activation and can suppress IL-2 production in conventional T cells. Thus, the control of IL-2 expression is critically important to T cell immune responses, yet the mechanisms remain incompletely understood. Nuclear factor (NF) 90 is a zinc-finger DNA- and double-stranded RNA-binding protein subunit that binds specifically to the antigen receptor response element (ARRE)/NF of activated T cells target sequence in the IL-2 proximal promoter. Inducible binding of NF90 to the IL-2 promoter in vivo is shown by chromatin immunoprecipitation. NF90 gene-targeted mice exhibit perinatal lethality. Compared with newborn NF90(+/+) mice, newborn NF90(-/-) mice demonstrate severe impairment of IL-2 expression. Compared with wild-type cells, T cells deficient in NF90 are impaired in ARRE and IL-2 transcriptional activation and IL-2 mRNA stabilization. Fetal liver cells from NF90 gene-targeted mice were transplanted into irradiated adult recombination activating gene (RAG)-2(-/-) and IL-2Rgamma(-/-) mice deficient in T cells, B cells, and natural killer cells. NF90(+/+)- and NF90(-/-)-RAG chimeric mice showed grossly normal repopulation of the thymus and spleen, but only NF90(-/-) T cells were severely impaired in IL-2 gene expression. Compared with littermates, NF90(-/-) RAG chimeric mice exhibited profound T cell lymphocytopenia in the peripheral circulation. Thus, NF90 regulates inducible IL-2 transcription, mRNA stability, and gene expression in T cells and represents a novel therapeutic target for the modulation of T cell immune responses. [Abstract/Link to Full Text]

Henry T, Brotcke A, Weiss DS, Thompson LJ, Monack DM
Type I interferon signaling is required for activation of the inflammasome during Francisella infection.
J Exp Med. 2007 May 14;204(5):987-94.
Francisella tularensis is a pathogenic bacterium whose virulence is linked to its ability to replicate within the host cell cytosol. Entry into the macrophage cytosol activates a host-protective multimolecular complex called the inflammasome to release the proinflammatory cytokines interleukin (IL)-1beta and -18 and trigger caspase-1-dependent cell death. In this study, we show that cytosolic F. tularensis subspecies novicida (F. novicida) induces a type I interferon (IFN) response that is essential for caspase-1 activation, inflammasome-mediated cell death, and release of IL-1beta and -18. Extensive type I IFN-dependent cell death resulting in macrophage depletion occurs in vivo during F. novicida infection. Type I IFN is also necessary for inflammasome activation in response to cytosolic Listeria monocytogenes but not vacuole-localized Salmonella enterica serovar Typhimurium or extracellular adenosine triphosphate. These results show the specific connection between type I IFN signaling and inflammasome activation, which are two sequential events triggered by the recognition of cytosolic bacteria. To our knowledge, this is the first example of the positive regulation of inflammasome activation. This connection underscores the importance of the cytosolic recognition of pathogens and highlights how multiple innate immunity pathways interact before commitment to critical host responses. [Abstract/Link to Full Text]

Summers-DeLuca LE, McCarthy DD, Cosovic B, Ward LA, Lo CC, Scheu S, Pfeffer K, Gommerman JL
Expression of lymphotoxin-alphabeta on antigen-specific T cells is required for DC function.
J Exp Med. 2007 May 14;204(5):1071-81.
During an immune response, activated antigen (Ag)-specific T cells condition dendritic cells (DCs) to enhance DC function and survival within the inflamed draining lymph node (LN). It has been difficult to ascertain the role of the tumor necrosis factor (TNF) superfamily member lymphotoxin-alphabeta (LTalphabeta) in this process because signaling through the LTbeta-receptor (LTbetaR) controls multiple aspects of lymphoid tissue organization. To resolve this, we have used an in vivo system where the expression of TNF family ligands is manipulated only on the Ag-specific T cells that interact with and condition Ag-bearing DCs. We report that LTalphabeta is a critical participant required for optimal DC function, independent of its described role in maintaining lymphoid tissue organization. In the absence of LTalphabeta or CD40L on Ag-specific T cells, DC dysfunction could be rescued in vivo via CD40 or LTbetaR stimulation, respectively, suggesting that these two pathways cooperate for optimal DC conditioning. [Abstract/Link to Full Text]

Boos MD, Yokota Y, Eberl G, Kee BL
Mature natural killer cell and lymphoid tissue-inducing cell development requires Id2-mediated suppression of E protein activity.
J Exp Med. 2007 May 14;204(5):1119-30.
The Id2 transcriptional repressor is essential for development of natural killer (NK) cells, lymphoid tissue-inducing (LTi) cells, and secondary lymphoid tissues. Id2 was proposed to regulate NK and LTi lineage specification from multipotent progenitors through suppression of E proteins. We report that NK cell progenitors are not reduced in the bone marrow (BM) of Id2(-/-) mice, demonstrating that Id2 is not essential for NK lineage specification. Rather, Id2 is required for development of mature (m) NK cells. We define the mechanism by which Id2 functions by showing that a reduction in E protein activity, through deletion of E2A, overcomes the need for Id2 in development of BM mNK cells, LTi cells, and secondary lymphoid tissues. However, mNK cells are not restored in the blood or spleen of Id2(-/-)E2A(-/-) mice, suggesting a role for Id2 in suppression of alternative E proteins after maturation. Interestingly, the few splenic mNK cells in Id2(-/-) and Id2(-/-)E2A(-/-) mice have characteristics of thymus-derived NK cells, which develop in the absence of Id2, implying a differential requirement for Id2 in BM and thymic mNK development. Our findings redefine the essential functions of Id2 in lymphoid development and provide insight into the dynamic regulation of E and Id proteins during this process. [Abstract/Link to Full Text]

Crouch EE, Li Z, Takizawa M, Fichtner-Feigl S, Gourzi P, Montańo C, Feigenbaum L, Wilson P, Janz S, Papavasiliou FN, Casellas R
Regulation of AID expression in the immune response.
J Exp Med. 2007 May 14;204(5):1145-56.
The B cell-specific enzyme activation-induced cytidine deaminase (AID) has been shown to be essential for isotype switching and affinity maturation of antibody genes during the immune response. Conversely, AID activity has also been linked to autoimmunity and tumorigenesis. Determining how AID expression is regulated in vivo is therefore central to understanding its role in health and disease. Here we use phylogenetic footprinting and high-resolution histone acetylation mapping to accurately demarcate AID gene regulatory boundaries. Based on this strategy, we identify a novel, positive regulatory element required for AID transcription. Furthermore, we generate two AID indicator mouse strains using bacterial artificial chromosomes that faithfully recapitulate endogenous AID expression. The first strain uses a green fluorescent protein reporter to identify B cells that actively express AID during the immune response. In the second strain, AID transcription affects the permanent expression of a yellow fluorescent protein reporter in post-germinal center and terminally differentiated lymphocytes. We demonstrate the usefulness of these novel strains by resolving recent contradictory observations on AID expression during B cell ontogeny. [Abstract/Link to Full Text]

Lühn K, Simmons CP, Moran E, Dung NT, Chau TN, Quyen NT, Thao le TT, Van Ngoc T, Dung NM, Wills B, Farrar J, McMichael AJ, Dong T, Rowland-Jones S
Increased frequencies of CD4+ CD25(high) regulatory T cells in acute dengue infection.
J Exp Med. 2007 May 14;204(5):979-85.
Dengue virus infection is an increasingly important tropical disease, causing 100 million cases each year. Symptoms range from mild febrile illness to severe hemorrhagic fever. The pathogenesis is incompletely understood, but immunopathology is thought to play a part, with antibody-dependent enhancement and massive immune activation of T cells and monocytes/macrophages leading to a disproportionate production of proinflammatory cytokines. We sought to investigate whether a defective population of regulatory T cells (T reg cells) could be contributing to immunopathology in severe dengue disease. CD4(+)CD25(high)FoxP3(+) T reg cells of patients with acute dengue infection of different severities showed a conventional phenotype. Unexpectedly, their capacity to suppress T cell proliferation and to secrete interleukin-10 was not altered. Moreover, T reg cells suppressed the production of vasoactive cytokines after dengue-specific stimulation. Furthermore, T reg cell frequencies and also T reg cell/effector T cell ratios were increased in patients with acute infection. A strong indication that a relative rise of T reg cell/effector T cell ratios is beneficial for disease outcome comes from patients with mild disease in which this ratio is significantly increased (P < 0.0001) in contrast to severe cases (P = 0.2145). We conclude that although T reg cells expand and function normally in acute dengue infection, their relative frequencies are insufficient to control the immunopathology of severe disease. [Abstract/Link to Full Text]

Soares H, Waechter H, Glaichenhaus N, Mougneau E, Yagita H, Mizenina O, Dudziak D, Nussenzweig MC, Steinman RM
A subset of dendritic cells induces CD4+ T cells to produce IFN-gamma by an IL-12-independent but CD70-dependent mechanism in vivo.
J Exp Med. 2007 May 14;204(5):1095-106.
Interferon (IFN)-gamma, a cytokine critical for resistance to infection and tumors, is produced by CD4(+) helper T lymphocytes after stimulation by cultured dendritic cells (DCs) that secrete a cofactor, interleukin (IL)-12. We have identified a major IL-12-independent pathway whereby DCs induce IFN-gamma-secreting T helper (Th)1 CD4(+) T cells in vivo. This pathway requires the membrane-associated tumor necrosis family member CD70 and was identified by targeting the LACK antigen from Leishmania major within an antibody to CD205 (DEC-205), an uptake receptor on a subset of DCs. Another major DC subset, targeted with 33D1 anti-DCIR2 antibody, also induced IFN-gamma in vivo but required IL-12, not CD70. Isolated CD205(+) DCs expressed cell surface CD70 when presenting antigen to T cell receptor transgenic T cells, and this distinction was independent of maturation stimuli. CD70 was also essential for CD205(+) DC function in vivo. Detection of the IL-12-independent IFN-gamma pathway was obscured with nontargeted LACK, which was presented by both DC subsets. This in situ analysis points to CD70 as a decision maker for Th1 differentiation by CD205(+) DCs, even in Th2-prone BALB/c animals and potentially in vaccine design. The results indicate that two DC subsets have innate propensities to differentially affect the Th1/Th2 balance in vivo and by distinct mechanisms. [Abstract/Link to Full Text]

Sood R, Zogg M, Westrick RJ, Guo YH, Kerschen EJ, Girardi G, Salmon JE, Coughlin SR, Weiler H
Fetal gene defects precipitate platelet-mediated pregnancy failure in factor V Leiden mothers.
J Exp Med. 2007 May 14;204(5):1049-56.
We describe a mouse model of fetal loss in factor V Leiden (FvL) mothers in which fetal loss is triggered when the maternal prothrombotic state coincides with fetal gene defects that reduce activation of the protein C anticoagulant pathway within the placenta. Fetal loss is caused by disruption of placental morphogenesis at the stage of labyrinth layer formation and occurs in the absence of overt placental thrombosis, infarction, or perfusion defects. Platelet depletion or elimination of protease-activated receptor 4 (Par4) from the mother allows normal placentation and prevents fetal loss. These findings establish a cause-effect relationship for the observed epidemiologic association between maternal FvL status and fetal loss and identify fetal gene defects as risk modifiers of pregnancy failure in prothrombotic mothers. Pregnancy failure is mediated by Par4-dependent activation of maternal platelets at the fetomaternal interface and likely involves a pathogenic pathway independent of occlusive thrombosis. Our results further demonstrate that the interaction of two given thrombosis risk factors produces markedly disparate consequences on disease manifestation (i.e., thrombosis or pregnancy loss), depending on the vascular bed in which this interaction occurs. [Abstract/Link to Full Text]

Aspord C, Pedroza-Gonzalez A, Gallegos M, Tindle S, Burton EC, Su D, Marches F, Banchereau J, Palucka AK
Breast cancer instructs dendritic cells to prime interleukin 13-secreting CD4+ T cells that facilitate tumor development.
J Exp Med. 2007 May 14;204(5):1037-47.
We previously reported (Bell, D., P. Chomarat, D. Broyles, G. Netto, G.M. Harb, S. Lebecque, J. Valladeau, J. Davoust, K.A. Palucka, and J. Banchereau. 1999. J. Exp. Med. 190: 1417-1426) that breast cancer tumors are infiltrated with mature dendritic cells (DCs), which cluster with CD4(+) T cells. We now show that CD4(+) T cells infiltrating breast cancer tumors secrete type 1 (interferon gamma) as well as high levels of type 2 (interleukin [IL] 4 and IL-13) cytokines. Immunofluorescence staining of tissue sections revealed intense IL-13 staining on breast cancer cells. The expression of phosphorylated signal transducer and activator of transcription 6 in breast cancer cells suggests that IL-13 actually delivers signals to cancer cells. To determine the link between breast cancer, DCs, and CD4(+) T cells, we implanted human breast cancer cell lines in nonobese diabetic/LtSz-scid/scid beta2 microglobulin-deficient mice engrafted with human CD34(+) hematopoietic progenitor cells and autologous T cells. There, CD4(+) T cells promote early tumor development. This is dependent on DCs and can be partially prevented by administration of IL-13 antagonists. Thus, breast cancer targets DCs to facilitate its development. [Abstract/Link to Full Text]

Esparza J, Yamada T
The discovery value of "Big Science".
J Exp Med. 2007 Apr 16;204(4):701-4.
The increasing complexity of biomedical research is leading to the exploration of new models for large-scale collaborative research. This Big Science approach, however, has created anxieties and potential tensions between investigator-driven research, and research guided by a more organized, collaborative effort. Another potential tension exists between research conducted purely in search of new knowledge and research aimed at finding solutions. We argue that big biomedicine--the work of coordinated multidisciplinary groups that use the latest technologies to solve complex problems--can be an important way to harness the creativity of individual investigators, stimulate innovation, and supply the infrastructure, experimental systems, and resources needed to solve the urgent health problems confronted by our global society. We discuss this using the example of the Global HIV Vaccine Enterprise. [Abstract/Link to Full Text]

Crozat K, Hoebe K, Ugolini S, Hong NA, Janssen E, Rutschmann S, Mudd S, Sovath S, Vivier E, Beutler B
Jinx, an MCMV susceptibility phenotype caused by disruption of Unc13d: a mouse model of type 3 familial hemophagocytic lymphohistiocytosis.
J Exp Med. 2007 Apr 16;204(4):853-63.
Mouse cytomegalovirus (MCMV) susceptibility often results from defects of natural killer (NK) cell function. Here we describe Jinx, an N-ethyl-N-nitrosourea-induced MCMV susceptibility mutation that permits unchecked proliferation of the virus, causing death. In Jinx homozygotes, activated NK cells and cytotoxic T lymphocytes (CTLs) fail to degranulate, although they retain the ability to produce cytokines, and cytokine levels are markedly elevated in the blood of infected mutant mice. Jinx was mapped to mouse chromosome 11 on a total of 246 meioses and confined to a 4.60-million basepair critical region encompassing 122 annotated genes. The phenotype was ascribed to the creation of a novel donor splice site in Unc13d, the mouse orthologue of human MUNC13-4, in which mutations cause type 3 familial hemophagocytic lymphohistiocytosis (FHL3), a fatal disease marked by massive hepatosplenomegaly, anemia, and thrombocytopenia. Jinx mice do not spontaneously develop clinical features of hemophagocytic lymphohistiocytosis (HLH), but do so when infected with lymphocytic choriomeningitis virus, exhibiting hyperactivation of CTLs and antigen-presenting cells, and inadequate restriction of viral proliferation. In contrast, neither Listeria monocytogenes nor MCMV induces the syndrome. In mice, the HLH phenotype is conditional, which suggests the existence of a specific infectious trigger of FHL3 in humans. [Abstract/Link to Full Text]

Ashton AW, Mukherjee S, Nagajyothi FN, Huang H, Braunstein VL, Desruisseaux MS, Factor SM, Lopez L, Berman JW, Wittner M, Scherer PE, Capra V, Coffman TM, Serhan CN, Gotlinger K, Wu KK, Weiss LM, Tanowitz HB
Thromboxane A2 is a key regulator of pathogenesis during Trypanosoma cruzi infection.
J Exp Med. 2007 Apr 16;204(4):929-40.
Chagas' disease is caused by infection with the parasite Trypanosoma cruzi. We report that infected, but not uninfected, human endothelial cells (ECs) released thromboxane A(2) (TXA(2)). Physical chromatography and liquid chromatography-tandem mass spectrometry revealed that TXA(2) is the predominant eicosanoid present in all life stages of T. cruzi. Parasite-derived TXA(2) accounts for up to 90% of the circulating levels of TXA(2) in infected wild-type mice, and perturbs host physiology. Mice in which the gene for the TXA(2) receptor (TP) has been deleted, exhibited higher mortality and more severe cardiac pathology and parasitism (fourfold) than WT mice after infection. Conversely, deletion of the TXA(2) synthase gene had no effect on survival or disease severity. TP expression on somatic cells, but not cells involved in either acquired or innate immunity, was the primary determinant of disease progression. The higher intracellular parasitism observed in TP-null ECs was ablated upon restoration of TP expression. We conclude that the host response to parasite-derived TXA(2) in T. cruzi infection is possibly an important determinant of mortality and parasitism. A deeper understanding of the role of TXA(2) may result in novel therapeutic targets for a disease with limited treatment options. [Abstract/Link to Full Text]

Waisman A, Kraus M, Seagal J, Ghosh S, Melamed D, Song J, Sasaki Y, Classen S, Lutz C, Brombacher F, Nitschke L, Rajewsky K
IgG1 B cell receptor signaling is inhibited by CD22 and promotes the development of B cells whose survival is less dependent on Ig alpha/beta.
J Exp Med. 2007 Apr 16;204(4):747-58.
We describe a mouse strain in which B cell development relies either on the expression of membrane-bound immunoglobulin (Ig) gamma1 or mu heavy chains. Progenitor cells expressing gamma1 chains from the beginning generate a peripheral B cell compartment of normal size with all subsets, but a partial block is seen at the pro- to pre-B cell transition. Accordingly, gamma1-driven B cell development is disfavored in competition with developing B cells expressing a wild-type (WT) IgH locus. However, the mutant B cells display a long half-life and accumulate in the mature B cell compartment, and even though partial truncation of the Ig alpha cytoplasmic tail compromises their development, it does not affect their maintenance, as it does in WT cells. IgG1-expressing B cells showed an enhanced Ca(2+) response upon B cell receptor cross-linking, which was not due to a lack of inhibition by CD22. The enhanced Ca(2+) response was also observed in mature B cells that had been switched from IgM to IgG1 expression in vivo. Collectively, these results suggest that the gamma1 chain can exert a unique signaling function that can partially replace that of the Ig alpha/beta heterodimer in B cell maintenance and may contribute to memory B cell physiology. [Abstract/Link to Full Text]

Shin H, Blackburn SD, Blattman JN, Wherry EJ
Viral antigen and extensive division maintain virus-specific CD8 T cells during chronic infection.
J Exp Med. 2007 Apr 16;204(4):941-9.
Efficient maintenance of memory CD8 T cells is central to long-term protective immunity. IL-7- and IL-15-driven homeostatic proliferation is essential for long-term memory CD8 T cell persistence after acute infections. During chronic infections, however, virus-specific CD8 T cells respond poorly to these cytokines. Yet, virus-specific CD8 T cells often persist for long periods of time during chronic infections. We have addressed this apparent paradox by examining the mechanism for maintaining virus-specific CD8 T cells during chronic infection. We find that homeostatic cytokines (e.g., IL-7/15), inflammatory signals, and priming of recent thymic emigrants are not sufficient to maintain virus-specific CD8 T cells over time during chronic infection. Rather, our results demonstrate that viral peptide is required for virus-specific CD8 T cell persistence during chronic infection. Moreover, this viral antigen-dependent maintenance results in a dramatically different type of T cell division than is normally observed during memory T cell homeostasis. Rather than undergoing slow, steady homeostatic turnover during chronic viral infection, CD8 T cells undergo extensive peptide-dependent division, yet cell numbers remain relatively stable. These results indicate that antigen-specific CD8 T cell responses during persisting infection are maintained by a mechanism distinct from that after acute infection. [Abstract/Link to Full Text]

Horikawa K, Martin SW, Pogue SL, Silver K, Peng K, Takatsu K, Goodnow CC
Enhancement and suppression of signaling by the conserved tail of IgG memory-type B cell antigen receptors.
J Exp Med. 2007 Apr 16;204(4):759-69.
Immunological memory is characterized by heightened immunoglobulin (Ig) G antibody production caused in part by enhanced plasma cell formation conferred by conserved transmembrane and cytoplasmic segments in isotype-switched IgG B cell receptors. We tested the hypothesis that the IgG tail enhances intracellular B cell antigen receptor (BCR) signaling responses to antigen by analyzing B cells from Ig transgenic mice with IgM receptors or chimeric IgMG receptors containing the IgG tail segment. The IgG tail segment enhanced intracellular calcium responses but not tyrosine or extracellular signal-related kinase (ERK) phosphorylation. Biochemical analysis and crosses to CD22-deficient mice established that IgG tail enhancement of calcium and antibody responses, as well as marginal zone B cell formation, was not due to diminished CD22 phosphorylation or inhibitory function. Microarray profiling showed no evidence for enhanced signaling by the IgG tail for calcium/calcineurin, ERK, or nuclear factor kappaB response genes and little evidence for any enhanced gene induction. Instead, almost half of the antigen-induced gene response in IgM B cells was diminished 50-90% by the IgG tail segment. These findings suggest a novel "less-is-more" hypothesis to explain how switching to IgG enhances B cell memory responses, whereby decreased BCR signaling to genes that oppose marginal zone and plasma cell differentiation enhances the formation of these key cell types. [Abstract/Link to Full Text]

Purton JF, Tan JT, Rubinstein MP, Kim DM, Sprent J, Surh CD
Antiviral CD4+ memory T cells are IL-15 dependent.
J Exp Med. 2007 Apr 16;204(4):951-61.
Survival and intermittent proliferation of memory CD4(+) and CD8(+) T cells appear to be controlled by different homeostatic mechanisms. In particular, contact with interleukin (IL)-15 has a decisive influence on memory CD8(+) cells, but not memory CD4(+) cells. Past studies of memory CD4(+) cells have relied heavily on the use of naturally occurring memory phenotype (MP) cells as a surrogate for antigen (Ag)-specific memory cells. However, we show here that MP CD4(+) cells contain a prominent subset of rapidly proliferating major histocompatibility complex (MHC) II-dependent cells. In contrast, Ag-specific memory CD4 cells have a slow turnover rate and are MHC II independent. In irradiated hosts, these latter cells ignore IL-15 and expand in response to the elevated levels of IL-7 in the lymphopenic hosts. In contrast, in normal nonlymphopenic hosts where IL-7 levels are low, memory CD4 cells are heavily dependent on IL-15. Significantly, memory CD4(+) responsiveness to endogenous IL-15 reflects marked competition from other cells, especially CD8(+) and natural killer cells, and increases considerably after removal of these cells. Therefore, under normal physiological conditions, homeostasis of CD8(+) and CD4(+) memory cells is quite similar and involves IL-15 and IL-7. [Abstract/Link to Full Text]

Tadokoro Y, Ema H, Okano M, Li E, Nakauchi H
De novo DNA methyltransferase is essential for self-renewal, but not for differentiation, in hematopoietic stem cells.
J Exp Med. 2007 Apr 16;204(4):715-22.
DNA methylation is an epigenetic modification essential for development. The DNA methyltransferases Dnmt3a and Dnmt3b execute de novo DNA methylation in gastrulating embryos and differentiating germline cells. It has been assumed that these enzymes generally play a role in regulating cell differentiation. To test this hypothesis, we examined the role of Dnmt3a and Dnmt3b in adult stem cells. CD34(-/low), c-Kit(+), Sca-1(+), lineage marker(-) (CD34(-) KSL) cells, a fraction of mouse bone marrow cells highly enriched in hematopoietic stem cells (HSCs), expressed both Dnmt3a and Dnmt3b. Using retroviral Cre gene transduction, we conditionally disrupted Dnmt3a, Dnmt3b, or both Dnmt3a and Dnmt3b (Dnmt3a/Dnmt3b) in CD34(-) KSL cells purified from mice in which the functional domains of these genes are flanked by two loxP sites. We found that Dnmt3a and Dnmt3b function as de novo DNA methyltransferases during differentiation of hematopoietic cells. Unexpectedly, in vitro colony assays and in vivo transplantation assays showed that both myeloid and lymphoid lineage differentiation potentials were maintained in Dnmt3a-, Dnmt3b-, and Dnmt3a/Dnmt3b-deficient HSCs. However, Dnmt3a/Dnmt3b-deficient HSCs, but not Dnmt3a- or Dnmt3b-deficient HSCs, were incapable of long-term reconstitution in transplantation assays. These findings establish a critical role for DNA methylation by Dnmt3a and Dnmt3b in HSC self-renewal. [Abstract/Link to Full Text]