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Recent Articles in PLoS Medicine / Public Libary of Science

Sripa B, Kaewkes S, Sithithaworn P, Mairiang E, Laha T, Smout M, Pairojkul C, Bhudhisawasdi V, Tesana S, Thinkamrop B, Bethony JM, Loukas A, Brindley PJ
Liver fluke induces cholangiocarcinoma.
PLoS Med. 2007 Jul;4(7):e201. [Abstract/Link to Full Text]

Riddell SR, Appelbaum FR
Graft-versus-host disease: a surge of developments.
PLoS Med. 2007 Jul;4(7):e198. [Abstract/Link to Full Text]

Killeen GF, Smith TA, Ferguson HM, Mshinda H, Abdulla S, Lengeler C, Kachur SP
Preventing childhood malaria in Africa by protecting adults from mosquitoes with insecticide-treated nets.
PLoS Med. 2007 Jul;4(7):e229.
BACKGROUND: Malaria prevention in Africa merits particular attention as the world strives toward a better life for the poorest. Insecticide-treated nets (ITNs) represent a practical means to prevent malaria in Africa, so scaling up coverage to at least 80% of young children and pregnant women by 2010 is integral to the Millennium Development Goals (MDG). Targeting individual protection to vulnerable groups is an accepted priority, but community-level impacts of broader population coverage are largely ignored even though they may be just as important. We therefore estimated coverage thresholds for entire populations at which individual- and community-level protection are equivalent, representing rational targets for ITN coverage beyond vulnerable groups. METHODS AND FINDINGS: Using field-parameterized malaria transmission models, we show that high (80% use) but exclusively targeted coverage of young children and pregnant women (representing <20% of the population) will deliver limited protection and equity for these vulnerable groups. In contrast, relatively modest coverage (35%-65% use, with this threshold depending on ecological scenario and net quality) of all adults and children, rather than just vulnerable groups, can achieve equitable community-wide benefits equivalent to or greater than personal protection. CONCLUSIONS: Coverage of entire populations will be required to accomplish large reductions of the malaria burden in Africa. While coverage of vulnerable groups should still be prioritized, the equitable and communal benefits of wide-scale ITN use by older children and adults should be explicitly promoted and evaluated by national malaria control programmes. ITN use by the majority of entire populations could protect all children in such communities, even those not actually covered by achieving existing personal protection targets of the MDG, Roll Back Malaria Partnership, or the US President's Malaria Initiative. [Abstract/Link to Full Text]

Mathenge W, Nkurikiye J, Limburg H, Kuper H
Rapid assessment of avoidable blindness in Western Rwanda: blindness in a postconflict setting.
PLoS Med. 2007 Jul;4(7):e217.
BACKGROUND: The World Health Organization estimates that there were 37 million blind people in 2002 and that the prevalence of blindness was 9% among adults in Africa aged 50 years or older. Recent surveys indicate that this figure may be overestimated, while a survey from southern Sudan suggested that postconflict areas are particularly vulnerable to blindness. The aim of this study was to conduct a Rapid Assessment for Avoidable Blindness to estimate the magnitude and causes of visual impairment in people aged > or = 50 y in the postconflict area of the Western Province of Rwanda, which includes one-quarter of the population of Rwanda. METHODS AND FINDINGS: Clusters of 50 people aged > or = 50 y were selected through probability proportionate to size sampling. Households within clusters were selected through compact segment sampling. Visual acuity (VA) was measured with a tumbling "E" chart, and those with VA below 6/18 in either eye were examined by an ophthalmologist. The teams examined 2,206 people (response rate 98.0%). The unadjusted prevalence of bilateral blindness was 1.8% (95% confidence interval [CI] 1.2%-2.4%), 1.3% (0.8%-1.7%) for severe visual impairment, and 5.3% (4.2%-6.4%) for visual impairment. Most bilateral blindness (65%) was due to cataract. Overall, the vast majority of cases of blindness (80.0%), severe visual impairment (67.9%), and visual impairment (87.2%) were avoidable (i.e.. due to cataract, refractive error, aphakia, trachoma, or corneal scar). The cataract surgical coverage was moderate; 47% of people with bilateral cataract blindness (VA < 3/60) had undergone surgery. Of the 29 eyes that had undergone cataract surgery, nine (31%) had a best-corrected poor outcome (i.e., VA < 6/60). Extrapolating these estimates to Rwanda's Western Province, among the people aged 50 years or above 2,565 are expected to be blind, 1,824 to have severe visual impairment, and 8,055 to have visual impairment. CONCLUSIONS: The prevalence of blindness and visual impairment in this postconflict area in the Western Province of Rwanda was far lower than expected. Most of the cases of blindness and visual impairment remain avoidable, however, suggesting that the implementation of an effective eye care service could reduce the prevalence further. [Abstract/Link to Full Text]

Gartner CE, Hall WD, Chapman S, Freeman B
Should the health community promote smokeless tobacco (snus) as a harm reduction measure?
PLoS Med. 2007 Jul;4(7):e185.
BACKGROUND TO THE DEBATE: The tobacco control community is divided on whether or not to inform the public that using oral, smokeless tobacco (Swedish snus) is less hazardous to health than smoking tobacco. Proponents of "harm reduction" point to the Swedish experience. Snus seems to be widely used as an alternative to cigarettes in Sweden, say these proponents, contributing to the low overall prevalence of smoking and smoking-related disease. Harm reduction proponents thus argue that the health community should actively inform inveterate cigarette smokers of the benefits of switching to snus. However, critics of harm reduction say that snus has its own risks, that no form of tobacco should ever be promoted, and that Sweden's experience is likely to be specific to that culture and not transferable to other settings. Critics also remain deeply suspicious that the tobacco industry will use snus marketing as a "gateway" to promote cigarettes. In the interests of promoting debate, the authors (who are collaborators on a research project on the future of tobacco control) have agreed to outline the strongest arguments for and against promoting Swedish snus as a form of harm reduction. [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]

Aguirre GK, Komáromy AM, Cideciyan AV, Brainard DH, Aleman TS, Roman AJ, Avants BB, Gee JC, Korczykowski M, Hauswirth WW, Acland GM, Aguirre GD, Jacobson SG
Canine and human visual cortex intact and responsive despite early retinal blindness from RPE65 mutation.
PLoS Med. 2007 Jun;4(6):e230.
BACKGROUND: RPE65 is an essential molecule in the retinoid-visual cycle, and RPE65 gene mutations cause the congenital human blindness known as Leber congenital amaurosis (LCA). Somatic gene therapy delivered to the retina of blind dogs with an RPE65 mutation dramatically restores retinal physiology and has sparked international interest in human treatment trials for this incurable disease. An unanswered question is how the visual cortex responds after prolonged sensory deprivation from retinal dysfunction. We therefore studied the cortex of RPE65-mutant dogs before and after retinal gene therapy. Then, we inquired whether there is visual pathway integrity and responsivity in adult humans with LCA due to RPE65 mutations (RPE65-LCA). METHODS AND FINDINGS: RPE65-mutant dogs were studied with fMRI. Prior to therapy, retinal and subcortical responses to light were markedly diminished, and there were minimal cortical responses within the primary visual areas of the lateral gyrus (activation amplitude mean +/- standard deviation [SD] = 0.07% +/- 0.06% and volume = 1.3 +/- 0.6 cm(3)). Following therapy, retinal and subcortical response restoration was accompanied by increased amplitude (0.18% +/- 0.06%) and volume (8.2 +/- 0.8 cm(3)) of activation within the lateral gyrus (p < 0.005 for both). Cortical recovery occurred rapidly (within a month of treatment) and was persistent (as long as 2.5 y after treatment). Recovery was present even when treatment was provided as late as 1-4 y of age. Human RPE65-LCA patients (ages 18-23 y) were studied with structural magnetic resonance imaging. Optic nerve diameter (3.2 +/- 0.5 mm) was within the normal range (3.2 +/- 0.3 mm), and occipital cortical white matter density as judged by voxel-based morphometry was slightly but significantly altered (1.3 SD below control average, p = 0.005). Functional magnetic resonance imaging in human RPE65-LCA patients revealed cortical responses with a markedly diminished activation volume (8.8 +/- 1.2 cm(3)) compared to controls (29.7 +/- 8.3 cm(3), p < 0.001) when stimulated with lower intensity light. Unexpectedly, cortical response volume (41.2 +/- 11.1 cm(3)) was comparable to normal (48.8 +/- 3.1 cm(3), p = 0.2) with higher intensity light stimulation. CONCLUSIONS: Visual cortical responses dramatically improve after retinal gene therapy in the canine model of RPE65-LCA. Human RPE65-LCA patients have preserved visual pathway anatomy and detectable cortical activation despite limited visual experience. Taken together, the results support the potential for human visual benefit from retinal therapies currently being aimed at restoring vision to the congenitally blind with genetic retinal disease. [Abstract/Link to Full Text]

Brogger J
Reporting of systematic reviews: better software required.
PLoS Med. 2007 Jun;4(6):e225. [Abstract/Link to Full Text]

Moher D, Tetzlaff J, Tricco AC, Sampson M, Altman DG
Epidemiology and reporting characteristics of systematic reviews.
PLoS Med. 2007 Mar 27;4(3):e78.
BACKGROUND: Systematic reviews (SRs) have become increasingly popular to a wide range of stakeholders. We set out to capture a representative cross-sectional sample of published SRs and examine them in terms of a broad range of epidemiological, descriptive, and reporting characteristics, including emerging aspects not previously examined. METHODS AND FINDINGS: We searched Medline for SRs indexed during November 2004 and written in English. Citations were screened and those meeting our inclusion criteria were retained. Data were collected using a 51-item data collection form designed to assess the epidemiological and reporting details and the bias-related aspects of the reviews. The data were analyzed descriptively. In total 300 SRs were identified, suggesting a current annual publication rate of about 2,500, involving more than 33,700 separate studies including one-third of a million participants. The majority (272 [90.7%]) of SRs were reported in specialty journals. Most reviews (213 [71.0%]) were categorized as therapeutic, and included a median of 16 studies involving 1,112 participants. Funding sources were not reported in more than one-third (122 [40.7%]) of the reviews. Reviews typically searched a median of three electronic databases and two other sources, although only about two-thirds (208 [69.3%]) of them reported the years searched. Most (197/295 [66.8%]) reviews reported information about quality assessment, while few (68/294 [23.1%]) reported assessing for publication bias. A little over half (161/300 [53.7%]) of the SRs reported combining their results statistically, of which most (147/161 [91.3%]) assessed for consistency across studies. Few (53 [17.7%]) SRs reported being updates of previously completed reviews. No review had a registration number. Only half (150 [50.0%]) of the reviews used the term "systematic review" or "meta-analysis" in the title or abstract. There were large differences between Cochrane reviews and non-Cochrane reviews in the quality of reporting several characteristics. CONCLUSIONS: SRs are now produced in large numbers, and our data suggest that the quality of their reporting is inconsistent. This situation might be improved if more widely agreed upon evidence-based reporting guidelines were endorsed and adhered to by authors and journals. These results substantiate the view that readers should not accept SRs uncritically. [Abstract/Link to Full Text]

Many reviews are systematic but some are more transparent and completely reported than others.
PLoS Med. 2007 Mar;4(3):e147. [Abstract/Link to Full Text]

The changing face of occupational medicine.
PLoS Med. 2007 Jun;4(6):e221. [Abstract/Link to Full Text]

Grudzen CR, Kerndt PR
The adult film industry: time to regulate?
PLoS Med. 2007 Jun;4(6):e126. [Abstract/Link to Full Text]

Villafuerte-Gálvez J, Curioso WH, Gayoso O
Biomedical journals and global poverty: is HINARI a step backwards?
PLoS Med. 2007 Jun;4(6):e220. [Abstract/Link to Full Text]

Ioannidis JP
Why most published research findings are false: author's reply to Goodman and Greenland.
PLoS Med. 2007 Jun;4(6):e215. [Abstract/Link to Full Text]

Goodman S, Greenland S
Why most published research findings are false: problems in the analysis.
PLoS Med. 2007 Apr;4(4):e168. [Abstract/Link to Full Text]

Reinhard R
Consent for genomic epidemiology in developing countries: added human subject protection also needed.
PLoS Med. 2007 Jun;4(6):e214. [Abstract/Link to Full Text]

Bonchek L
Lethal injection: let's be honest about the death penalty.
PLoS Med. 2007 Jun;4(6):e213; discussion e224. [Abstract/Link to Full Text]

Bermejo-Martin JF, Kelvin DJ, Guan Y, Chen H, Perez-Breńa P, Casas I, Arranz E, de Lejarazu RO
Neuraminidase antibodies and H5N1: geographic-dependent influenza epidemiology could determine cross-protection against emerging strains.
PLoS Med. 2007 Jun;4(6):e212. [Abstract/Link to Full Text]

Sandbulte MR, Jimenez GS, Boon AC, Smith LR, Treanor JJ, Webby RJ
Cross-reactive neuraminidase antibodies afford partial protection against H5N1 in mice and are present in unexposed humans.
PLoS Med. 2007 Feb;4(2):e59.
BACKGROUND: A pandemic H5N1 influenza outbreak would be facilitated by an absence of immunity to the avian-derived virus in the human population. Although this condition is likely in regard to hemagglutinin-mediated immunity, the neuraminidase (NA) of H5N1 viruses (avN1) and of endemic human H1N1 viruses (huN1) are classified in the same serotype. We hypothesized that an immune response to huN1 could mediate cross-protection against H5N1 influenza virus infection. METHODS AND FINDINGS: Mice were immunized against the NA of a contemporary human H1N1 strain by DNA vaccination. They were challenged with recombinant A/Puerto Rico/8/34 (PR8) viruses bearing huN1 (PR8-huN1) or avN1 (PR8-avN1) or with H5N1 virus A/Vietnam/1203/04. Additional naďve mice were injected with sera from vaccinated mice prior to H5N1 challenge. Also, serum specimens from humans were analyzed for reactivity with avN1. Immunization elicited a serum IgG response to huN1 and robust protection against the homologous challenge virus. Immunized mice were partially protected from lethal challenge with H5N1 virus or recombinant PR8-avN1. Sera transferred from immunized mice to naďve animals conferred similar protection against H5N1 mortality. Analysis of human sera showed that antibodies able to inhibit the sialidase activity of avN1 exist in some individuals. CONCLUSIONS: These data reveal that humoral immunity elicited by huN1 can partially protect against H5N1 infection in a mammalian host. Our results suggest that a portion of the human population could have some degree of resistance to H5N1 influenza, with the possibility that this could be induced or enhanced through immunization with seasonal influenza vaccines. [Abstract/Link to Full Text]

Khoury MJ, Little J, Higgins J, Ioannidis JP, Gwinn M
Reporting of systematic reviews: the challenge of genetic association studies.
PLoS Med. 2007 Jun;4(6):e211. [Abstract/Link to Full Text]

Reis BY, Kohane IS, Mandl KD
An epidemiological network model for disease outbreak detection.
PLoS Med. 2007 Jun;4(6):e210.
BACKGROUND: Advanced disease-surveillance systems have been deployed worldwide to provide early detection of infectious disease outbreaks and bioterrorist attacks. New methods that improve the overall detection capabilities of these systems can have a broad practical impact. Furthermore, most current generation surveillance systems are vulnerable to dramatic and unpredictable shifts in the health-care data that they monitor. These shifts can occur during major public events, such as the Olympics, as a result of population surges and public closures. Shifts can also occur during epidemics and pandemics as a result of quarantines, the worried-well flooding emergency departments or, conversely, the public staying away from hospitals for fear of nosocomial infection. Most surveillance systems are not robust to such shifts in health-care utilization, either because they do not adjust baselines and alert-thresholds to new utilization levels, or because the utilization shifts themselves may trigger an alarm. As a result, public-health crises and major public events threaten to undermine health-surveillance systems at the very times they are needed most. METHODS AND FINDINGS: To address this challenge, we introduce a class of epidemiological network models that monitor the relationships among different health-care data streams instead of monitoring the data streams themselves. By extracting the extra information present in the relationships between the data streams, these models have the potential to improve the detection capabilities of a system. Furthermore, the models' relational nature has the potential to increase a system's robustness to unpredictable baseline shifts. We implemented these models and evaluated their effectiveness using historical emergency department data from five hospitals in a single metropolitan area, recorded over a period of 4.5 y by the Automated Epidemiological Geotemporal Integrated Surveillance real-time public health-surveillance system, developed by the Children's Hospital Informatics Program at the Harvard-MIT Division of Health Sciences and Technology on behalf of the Massachusetts Department of Public Health. We performed experiments with semi-synthetic outbreaks of different magnitudes and simulated baseline shifts of different types and magnitudes. The results show that the network models provide better detection of localized outbreaks, and greater robustness to unpredictable shifts than a reference time-series modeling approach. CONCLUSIONS: The integrated network models of epidemiological data streams and their interrelationships have the potential to improve current surveillance efforts, providing better localized outbreak detection under normal circumstances, as well as more robust performance in the face of shifts in health-care utilization during epidemics and major public events. [Abstract/Link to Full Text]

Wilson D, Ford N, Ngammee V, Chua A, Kyaw MK
HIV prevention, care, and treatment in two prisons in Thailand.
PLoS Med. 2007 Jun;4(6):e204. [Abstract/Link to Full Text]

Lagarde E
Road traffic injury is an escalating burden in Africa and deserves proportionate research efforts.
PLoS Med. 2007 Jun;4(6):e170. [Abstract/Link to Full Text]

Fraser C
Influenza pandemic vaccines: spread them thin?
PLoS Med. 2007 Jun;4(6):e228. [Abstract/Link to Full Text]

Riley S, Wu JT, Leung GM
Optimizing the dose of pre-pandemic influenza vaccines to reduce the infection attack rate.
PLoS Med. 2007 Jun;4(6):e218.
BACKGROUND: The recent spread of avian influenza in wild birds and poultry may be a precursor to the emergence of a 1918-like human pandemic. Therefore, stockpiles of human pre-pandemic vaccine (targeted at avian strains) are being considered. For many countries, the principal constraint for these vaccine stockpiles will be the total mass of antigen maintained. We tested the hypothesis that lower individual doses (i.e., less than the recommended dose for maximum protection) may provide substantial extra community-level benefits because they would permit wider vaccine coverage for a given total size of antigen stockpile. METHODS AND FINDINGS: We used a mathematical model to predict infection attack rates under different policies. The model incorporated both an individual's response to vaccination at different doses and the process of person-to-person transmission of pandemic influenza. We found that substantial reductions in the attack rate are likely if vaccines are given to more people at lower doses. These results are applicable to all three vaccine candidates for which data are available. As a guide to the magnitude of the effect, we simulated epidemics based on historical studies of immunogenicity. For example, for one of the vaccines for which data are available, the attack rate would drop from 67.6% to 58.7% if 160 out of the total US population of 300 million were given an optimal dose rather than 20 out of 300 million given the maximally protective dose (as promulgated in the US National Pandemic Preparedness Plan). Our results are conservative with respect to a number of alternative assumptions about the precise nature of vaccine protection. We also considered a model variant that includes a single high-risk subgroup representing children. For smaller stockpile sizes that allow vaccine to be offered only to the high-risk group at the optimal dose, the predicted benefits of using the homogenous model formed a lower bound in the presence of a risk group, even when the high-risk group was twice as infective and twice as susceptible. CONCLUSIONS: In addition to individual-level protection (i.e., vaccine efficacy), the population-level implications of pre-pandemic vaccine programs should be considered when deciding on stockpile size and dose. Our results suggest that a lower vaccine dose may be justified in order to increase population coverage, thereby reducing the infection attack rate overall. [Abstract/Link to Full Text]

Sievänen H, Kannus P
Physical activity reduces the risk of fragility fracture.
PLoS Med. 2007 Jun;4(6):e222. [Abstract/Link to Full Text]

Michaëlsson K, Olofsson H, Jensevik K, Larsson S, Mallmin H, Berglund L, Vessby B, Melhus H
Leisure physical activity and the risk of fracture in men.
PLoS Med. 2007 Jun;4(6):e199.
BACKGROUND: Data from previous studies are inconsistent, and it is therefore uncertain whether, to what extent, and at what level leisure physical activity influences the risk of osteoporotic fractures in men. METHODS AND FINDINGS: A cohort of 2,205 men, 49-51 y of age, was enrolled in a longitudinal, population-based study. Leisure physical activity and other lifestyle habits were established at baseline and at ages 60, 70, 77, and 82 y. During 35 y of follow-up, 482 men had at least one fracture. Cox's proportional hazards regression was used to determine hazard ratios (HRs) of fracture associated with time-dependent physical activity habits and covariates. Men with a sedentary lifestyle (HR 2.56, 95% confidence interval 1.55-4.24) or men who walked or bicycled only for pleasure (HR 1.61, 95% confidence interval 1.10-2.36) had an increased adjusted risk of hip fracture compared with men who participated in regular sports activities for at least 3 h/wk. At the end of follow-up, 8.4% of the men with a high physical activity, 13.3% of the men with a medium physical activity, and 20.5% of the men with a low physical activity had suffered a hip fracture. According to the estimation of population-attributable risk, one third of all hip fractures could be prevented by participation in regular sports activities. High activity also conferred a reduced overall fracture risk. CONCLUSIONS: Our data indicate that regular sports activities can reduce the risk of fractures in older men. [Abstract/Link to Full Text]

Kuntz AN, Davioud-Charvet E, Sayed AA, Califf LL, Dessolin J, Arnér ES, Williams DL
Thioredoxin glutathione reductase from Schistosoma mansoni: an essential parasite enzyme and a key drug target.
PLoS Med. 2007 Jun;4(6):e206.
BACKGROUND: Schistosomiasis--infection with helminth parasites in the genus Schistosoma, including S. mansoni--is a widespread, devastating tropical disease affecting more than 200 million people. No vaccine is available, and praziquantel, the only drug extensively utilized, is currently administered more than 100 million people yearly. Because praziquantel resistance may develop it is essential to identify novel drug targets. Our goal was to investigate the potential of a unique, selenium-containing parasite enzyme thioredoxin glutathione reductase (TGR) as a drug target. METHODS AND FINDINGS: Using RNA interference we found that TGR is essential for parasite survival; after silencing of TGR expression, in vitro parasites died within 4 d. We also found that auranofin is an efficient inhibitor of pure TGR (Ki = 10 nM), able to kill parasites rapidly in culture at physiological concentrations (5 microM), and able to partially cure infected mice (worm burden reductions of ~60%). Furthermore, two previously used antischistosomal compounds inhibited TGR activity, suggesting that TGR is a key target during therapy with those compounds. CONCLUSIONS: Collectively, our results indicate that parasite TGR meets all the major criteria to be a key target for antischistosomal chemotherapy. To our knowledge this is the first validation of a Schistosoma drug target using a convergence of both genetic and biochemical approaches. [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]

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]

Lyssenko V, Almgren P, Anevski D, Orho-Melander M, Sjögren M, Saloranta C, Tuomi T, Groop L
Genetic prediction of future type 2 diabetes.
PLoS Med. 2005 Nov 1;2(12):e345.
BACKGROUND: Type 2 diabetes (T2D) is a multifactorial disease in which environmental triggers interact with genetic variants in the predisposition to the disease. A number of common variants have been associated with T2D but our knowledge of their ability to predict T2D prospectively is limited. METHODS AND FINDINGS: By using a Cox proportional hazard model, common variants in the PPARG (P12A), CAPN10 (SNP43 and 44), KCNJ11 (E23K), UCP2 (-866G>A), and IRS1 (G972R) genes were studied for their ability to predict T2D in 2,293 individuals participating in the Botnia study in Finland. After a median follow-up of 6 y, 132 (6%) persons developed T2D. The hazard ratio for risk of developing T2D was 1.7 (95% confidence interval [CI] 1.1-2.7) for the PPARG PP genotype, 1.5 (95% CI 1.0-2.2) for the CAPN10 SNP44 TT genotype, and 2.6 (95% CI 1.5-4.5) for the combination of PPARG and CAPN10 risk genotypes. In individuals with fasting plasma glucose > or = 5.6 mmol/l and body mass index > or = 30 kg/m(2), the hazard ratio increased to 21.2 (95% CI 8.7-51.4) for the combination of the PPARG PP and CAPN10 SNP43/44 GG/TT genotypes as compared to those with the low-risk genotypes with normal fasting plasma glucose and body mass index < 30 kg/m(2). CONCLUSION: We demonstrate in a large prospective study that variants in the PPARG and CAPN10 genes predict future T2D. Genetic testing might become a future approach to identify individuals at risk of developing T2D. [Abstract/Link to Full Text]

Lethal injection is not humane.
PLoS Med. 2007 Apr;4(4):e171. [Abstract/Link to Full Text]

Zimmers TA, Sheldon J, Lubarsky DA, López-Muńoz F, Waterman L, Weisman R, Koniaris LG
Lethal injection for execution: chemical asphyxiation?
PLoS Med. 2007 Apr;4(4):e156.
BACKGROUND: Lethal injection for execution was conceived as a comparatively humane alternative to electrocution or cyanide gas. The current protocols are based on one improvised by a medical examiner and an anesthesiologist in Oklahoma and are practiced on an ad hoc basis at the discretion of prison personnel. Each drug used, the ultrashort-acting barbiturate thiopental, the neuromuscular blocker pancuronium bromide, and the electrolyte potassium chloride, was expected to be lethal alone, while the combination was intended to produce anesthesia then death due to respiratory and cardiac arrest. We sought to determine whether the current drug regimen results in death in the manner intended. METHODS AND FINDINGS: We analyzed data from two US states that release information on executions, North Carolina and California, as well as the published clinical, laboratory, and veterinary animal experience. Execution outcomes from North Carolina and California together with interspecies dosage scaling of thiopental effects suggest that in the current practice of lethal injection, thiopental might not be fatal and might be insufficient to induce surgical anesthesia for the duration of the execution. Furthermore, evidence from North Carolina, California, and Virginia indicates that potassium chloride in lethal injection does not reliably induce cardiac arrest. CONCLUSIONS: We were able to analyze only a limited number of executions. However, our findings suggest that current lethal injection protocols may not reliably effect death through the mechanisms intended, indicating a failure of design and implementation. If thiopental and potassium chloride fail to cause anesthesia and cardiac arrest, potentially aware inmates could die through pancuronium-induced asphyxiation. Thus the conventional view of lethal injection leading to an invariably peaceful and painless death is questionable. [Abstract/Link to Full Text]

PLoS Medicine's Advisory Group on Publication Ethics.
PLoS Med. 2007 Feb;4(2):e81. [Abstract/Link to Full Text]

Coleman K, Hamblin R
Can pay-for-performance improve quality and reduce health disparities?
PLoS Med. 2007 Jun;4(6):e216. [Abstract/Link to Full Text]

Millett C, Gray J, Saxena S, Netuveli G, Khunti K, Majeed A
Ethnic disparities in diabetes management and pay-for-performance in the UK: the Wandsworth Prospective Diabetes Study.
PLoS Med. 2007 Jun;4(6):e191.
BACKGROUND: Pay-for-performance rewards health-care providers by paying them more if they succeed in meeting performance targets. A new contract for general practitioners in the United Kingdom represents the most radical shift towards pay-for-performance seen in any health-care system. The contract provides an important opportunity to address disparities in chronic disease management between ethnic and socioeconomic groups. We examined disparities in management of people with diabetes and intermediate clinical outcomes within a multiethnic population in primary care before and after the introduction of the new contract in April 2004. METHODS AND FINDINGS: We conducted a population-based longitudinal survey, using electronic general practice records, in an ethnically diverse part of southwest London. Outcome measures were prescribing levels and achievement of national treatment targets (HbA1c < or = 7.0%; blood pressure [BP] < 140/80 mm Hg; total cholesterol < or = 5 mmol/l or 193 mg/dl). The proportion of patients reaching treatment targets for HbA1c, BP, and total cholesterol increased significantly after the implementation of the new contract. The extents of these increases were broadly uniform across ethnic groups, with the exception of the black Caribbean patient group, which had a significantly lower improvement in HbA1c (adjusted odds ratio [AOR] 0.75, 95% confidence interval [CI] 0.57-0.97) and BP control (AOR 0.65, 95% CI 0.53-0.81) relative to the white British patient group. Variations in prescribing and achievement of treatment targets between ethnic groups present in 2003 were not attenuated in 2005. CONCLUSIONS: Pay-for-performance incentives have not addressed disparities in the management and control of diabetes between ethnic groups. Quality improvement initiatives must place greater emphasis on minority communities to avoid continued disparities in mortality from cardiovascular disease and the other major complications of diabetes. [Abstract/Link to Full Text]

Pai M, O'Brien R
Serial testing for tuberculosis: can we make sense of T cell assay conversions and reversions?
PLoS Med. 2007 Jun;4(6):e208. [Abstract/Link to Full Text]

Hill PC, Brookes RH, Fox A, Jackson-Sillah D, Jeffries DJ, Lugos MD, Donkor SA, Adetifa IM, de Jong BC, Aiken AM, Adegbola RA, McAdam KP
Longitudinal assessment of an ELISPOT test for Mycobacterium tuberculosis infection.
PLoS Med. 2007 Jun;4(6):e192.
BACKGROUND: Very little longitudinal information is available regarding the performance of T cell-based tests for Mycobacterium tuberculosis infection. To address this deficiency, we conducted a longitudinal assessment of the enzyme-linked immunosorbent spot test (ELISPOT) test in comparison to the standard tuberculin skin test (TST). METHODS AND FINDINGS: In tuberculosis (TB) contacts we repeated ELISPOT tests 3 mo (n = 341) and 18 mo (n = 210) after recruitment and TSTs at 18 mo (n = 130). We evaluated factors for association with conversion and reversion and investigated suspected cases of TB. Of 207 ELISPOT-negative contacts, 51 (24.6%) had 3-mo ELISPOT conversion, which was associated with a positive recruitment TST (odds ratio [OR] 2.2, 95% confidence interval [CI] 1.0-5.0, p = 0.048) and negatively associated with bacillus Calmette-Guérin (BCG) vaccination (OR 0.5, 95% CI 0.2-1.0, p = 0.06). Of 134 contacts, 54 (40.2%) underwent 3-mo ELISPOT reversion, which was less likely in those with a positive recruitment TST (OR 0.3, 95% CI 0.1-0.8, p = 0.014). Between 3 and 18 mo, 35/132 (26.5%) contacts underwent ELISPOT conversion and 28/78 (35.9%) underwent ELISPOT reversion. Of the 210 contacts with complete results, 73 (34.8%) were ELISPOT negative at all three time points; 36 (17.1%) were positive at all three time points. Between recruitment and 18 mo, 20 (27%) contacts had ELISPOT conversion; 37 (50%) had TST conversion, which was associated with a positive recruitment ELISPOT (OR 7.2, 95% CI 1.4-37.1, p = 0.019); 18 (32.7%) underwent ELISPOT reversion; and five (8.9%) underwent TST reversion. Results in 13 contacts diagnosed as having TB were mixed, but suggested higher TST sensitivity. CONCLUSIONS: Both ELISPOT conversion and reversion occur after M. tuberculosis exposure. Rapid ELISPOT reversion may reflect M. tuberculosis clearance or transition into dormancy and may contribute to the relatively low reported ELISPOT conversion rate. Therefore, a negative ELISPOT test for M. tuberculosis infection should be interpreted with caution. [Abstract/Link to Full Text]

Steingart KR, Henry M, Laal S, Hopewell PC, Ramsay A, Menzies D, Cunningham J, Weldingh K, Pai M
Commercial serological antibody detection tests for the diagnosis of pulmonary tuberculosis: a systematic review.
PLoS Med. 2007 Jun;4(6):e202.
BACKGROUND: The global tuberculosis epidemic results in nearly 2 million deaths and 9 million new cases of the disease a year. The vast majority of tuberculosis patients live in developing countries, where the diagnosis of tuberculosis relies on the identification of acid-fast bacilli on unprocessed sputum smears using conventional light microscopy. Microscopy has high specificity in tuberculosis-endemic countries, but modest sensitivity which varies among laboratories (range 20% to 80%). Moreover, the sensitivity is poor for paucibacillary disease (e.g., pediatric and HIV-associated tuberculosis). Thus, the development of rapid and accurate new diagnostic tools is imperative. Immune-based tests are potentially suitable for use in low-income countries as some test formats can be performed at the point of care without laboratory equipment. Currently, dozens of distinct commercial antibody detection tests are sold in developing countries. The question is "do they work?" METHODS AND FINDINGS: We conducted a systematic review to assess the accuracy of commercial antibody detection tests for the diagnosis of pulmonary tuberculosis. Studies from all countries using culture and/or microscopy smear for confirmation of pulmonary tuberculosis were eligible. Studies with fewer than 50 participants (25 patients and 25 control participants) were excluded. In a comprehensive search, we identified 68 studies. The results demonstrate that (1) overall, commercial tests vary widely in performance; (2) sensitivity is higher in smear-positive than smear-negative samples; (3) in studies of smear-positive patients, Anda-TB IgG by enzyme-linked immunosorbent assay shows limited sensitivity (range 63% to 85%) and inconsistent specificity (range 73% to 100%); (4) specificity is higher in healthy volunteers than in patients in whom tuberculosis disease is initially suspected and subsequently ruled out; and (5) there are insufficient data to determine the accuracy of most commercial tests in smear microscopy-negative patients, as well as their performance in children or persons with HIV infection. CONCLUSIONS: None of the commercial tests evaluated perform well enough to replace sputum smear microscopy. Thus, these tests have little or no role in the diagnosis of pulmonary tuberculosis. Lack of methodological rigor in these studies was identified as a concern. It will be important to review the basic science literature evaluating serological tests for the diagnosis of pulmonary tuberculosis to determine whether useful antigens have been described but their potential has not been fully exploited. Activities leading to the discovery of new antigens with immunodiagnostic potential need to be intensified. [Abstract/Link to Full Text]

Valdiserri RO
Late HIV diagnosis: bad medicine and worse public health.
PLoS Med. 2007 Jun;4(6):e200. [Abstract/Link to Full Text]

Holtgrave DR
Costs and consequences of the US Centers for Disease Control and Prevention's recommendations for opt-out HIV testing.
PLoS Med. 2007 Jun;4(6):e194.
BACKGROUND: The United States Centers for Disease Control and Prevention (CDC) recently recommended opt-out HIV testing (testing without the need for risk assessment and counseling) in all health care encounters in the US for persons 13-64 years old. However, the overall costs and consequences of these recommendations have not been estimated before. In this paper, I estimate the costs and public health impact of opt-out HIV testing relative to testing accompanied by client-centered counseling, and relative to a more targeted counseling and testing strategy. METHODS AND FINDINGS: Basic methods of scenario and cost-effectiveness analysis were used, from a payer's perspective over a one-year time horizon. I found that for the same programmatic cost of US$864,207,288, targeted counseling and testing services (at a 1% HIV seropositivity rate) would be preferred to opt-out testing: targeted services would newly diagnose more HIV infections (188,170 versus 56,940), prevent more HIV infections (14,553 versus 3,644), and do so at a lower gross cost per infection averted (US$59,383 versus US$237,149). While the study is limited by uncertainty in some input parameter values, the findings were robust across a variety of assumptions about these parameter values (including the estimated HIV seropositivity rate in the targeted counseling and testing scenario). CONCLUSIONS: While opt-out testing may be able to newly diagnose over 56,000 persons living with HIV in one year, abandoning client-centered counseling has real public health consequences in terms of HIV infections that could have been averted. Further, my analyses indicate that even when HIV seropositivity rates are as low as 0.3%, targeted counseling and testing performs better than opt-out testing on several key outcome variables. These analytic findings should be kept in mind as HIV counseling and testing policies are debated in the US. [Abstract/Link to Full Text]

Lee TH, Seng S, Sekine M, Hinton C, Fu Y, Avraham HK, Avraham S
Vascular endothelial growth factor mediates intracrine survival in human breast carcinoma cells through internally expressed VEGFR1/FLT1.
PLoS Med. 2007 Jun;4(6):e186.
BACKGROUND: While vascular endothelial growth factor (VEGF) expression in breast tumors has been correlated with a poor outcome in the pathogenesis of breast cancer, the expression, localization, and function of VEGF receptors VEGFR1 (also known as FLT1) and VEGFR2 (also known as KDR or FLK1), as well as neuropilin 1 (NRP1), in breast cancer are controversial. METHODS AND FINDINGS: We investigated the expression and function of VEGF and VEGF receptors in breast cancer cells. We observed that VEGFR1 expression was abundant, VEGFR2 expression was low, and NRP1 expression was variable. MDA-MB-231 and MCF-7 breast cancer cells, transfected with antisense VEGF cDNA or with siVEGF (VEGF-targeted small interfering RNA), showed a significant reduction in VEGF expression and increased apoptosis as compared to the control cells. Additionally, specifically targeted knockdown of VEGFR1 expression by siRNA (siVEGFR1) significantly decreased the survival of breast cancer cells through down-regulation of protein kinase B (AKT) phosphorylation, while targeted knockdown of VEGFR2 or NRP1 expression had no effect on the survival of these cancer cells. Since a VEGFR1-specific ligand, placenta growth factor (PGF), did not, as expected, inhibit the breast cancer cell apoptosis induced by siVEGF, and since VEGFR1 antibody also had no effects on the survival of these cells, we examined VEGFR1 localization. VEGFR1 was predominantly expressed internally in MDA-MB-231 and MCF-7 breast cancer cells. Specifically, VEGFR1 was found to be colocalized with lamin A/C and was expressed mainly in the nuclear envelope in breast cancer cell lines and primary breast cancer tumors. Breast cancer cells treated with siVEGFR1 showed significantly decreased VEGFR1 expression levels and a lack of VEGFR1 expression in the nuclear envelope. CONCLUSIONS: This study provides, to our knowledge for the first time, evidence of a unique survival system in breast cancer cells by which VEGF can act as an internal autocrine (intracrine) survival factor through its binding to VEGFR1. These results may lead to an improved strategy for tumor therapy based on the inhibition of angiogenesis. [Abstract/Link to Full Text]

Bero L, Oostvogel F, Bacchetti P, Lee K
Factors associated with findings of published trials of drug-drug comparisons: why some statins appear more efficacious than others.
PLoS Med. 2007 Jun;4(6):e184.
BACKGROUND: Published pharmaceutical industry-sponsored trials are more likely than non-industry-sponsored trials to report results and conclusions that favor drug over placebo. Little is known about potential biases in drug-drug comparisons. This study examined associations between research funding source, study design characteristics aimed at reducing bias, and other factors that potentially influence results and conclusions in randomized controlled trials (RCTs) of statin-drug comparisons. METHODS AND FINDINGS: This is a cross-sectional study of 192 published RCTs comparing a statin drug to another statin drug or non-statin drug. Data on concealment of allocation, selection bias, blinding, sample size, disclosed funding source, financial ties of authors, results for primary outcomes, and author conclusions were extracted by two coders (weighted kappa 0.80 to 0.97). Univariate and multivariate logistic regression identified associations between independent variables and favorable results and conclusions. Of the RCTs, 50% (95/192) were funded by industry, and 37% (70/192) did not disclose any funding source. Looking at the totality of available evidence, we found that almost all studies (98%, 189/192) used only surrogate outcome measures. Moreover, study design weaknesses common to published statin-drug comparisons included inadequate blinding, lack of concealment of allocation, poor follow-up, and lack of intention-to-treat analyses. In multivariate analysis of the full sample, trials with adequate blinding were less likely to report results favoring the test drug, and sample size was associated with favorable conclusions when controlling for other factors. In multivariate analysis of industry-funded RCTs, funding from the test drug company was associated with results (odds ratio = 20.16 [95% confidence interval 4.37-92.98], p < 0.001) and conclusions (odds ratio = 34.55 [95% confidence interval 7.09-168.4], p < 0.001) that favor the test drug when controlling for other factors. Studies with adequate blinding were less likely to report statistically significant results favoring the test drug. CONCLUSIONS: RCTs of head-to-head comparisons of statins with other drugs are more likely to report results and conclusions favoring the sponsor's product compared to the comparator drug. This bias in drug-drug comparison trials should be considered when making decisions regarding drug choice. [Abstract/Link to Full Text]

Villafuerte-Galvez J, Curioso WH
Teaching global health at the frontlines. A multidisciplinary course in Peru presents basic concepts to students.
PLoS Med. 2007 Jun;4(6):e130. [Abstract/Link to Full Text]

Kouyaté B, Sie A, Yé M, De Allegri M, Müller O
The great failure of malaria control in Africa: a district perspective from Burkina Faso.
PLoS Med. 2007 Jun;4(6):e127. [Abstract/Link to Full Text]

How is WHO responding to global public health threats?
PLoS Med. 2007 May;4(5):e197. [Abstract/Link to Full Text]

Schünemann HJ, Hill SR, Kakad M, Vist GE, Bellamy R, Stockman L, Wislřff TF, Del Mar C, Hayden F, Uyeki TM, Farrar J, Yazdanpanah Y, Zucker H, Beigel J, Chotpitayasunondh T, Hien TT, Ozbay B, Sugaya N, Oxman AD
Transparent development of the WHO rapid advice guidelines.
PLoS Med. 2007 May;4(5):e119.
Emerging health problems require rapid advice. We describe the development and pilot testing of a systematic, transparent approach used by the World Health Organization (WHO) to develop rapid advice guidelines in response to requests from member states confronted with uncertainty about the pharmacological management of avian influenza A (H5N1) virus infection. We first searched for systematic reviews of randomized trials of treatment and prevention of seasonal influenza and for non-trial evidence on H5N1 infection, including case reports and animal and in vitro studies. A panel of clinical experts, clinicians with experience in treating patients with H5N1, influenza researchers, and methodologists was convened for a two-day meeting. Panel members reviewed the evidence prior to the meeting and agreed on the process. It took one month to put together a team to prepare the evidence profiles (i.e., summaries of the evidence on important clinical and policy questions), and it took the team only five weeks to prepare and revise the evidence profiles and to prepare draft guidelines prior to the panel meeting. A draft manuscript for publication was prepared within 10 days following the panel meeting. Strengths of the process include its transparency and the short amount of time used to prepare these WHO guidelines. The process could be improved by shortening the time required to commission evidence profiles. Further development is needed to facilitate stakeholder involvement, and evaluate and ensure the guideline's usefulness. [Abstract/Link to Full Text]

Wilson K, Gardam M, McDougall C, Attaran A, Upshur R
WHO's response to global public health threats: XDR-TB.
PLoS Med. 2007 Jul 31;4(7):e246. [Abstract/Link to Full Text]

Levin H
Natural ventilation for prevention of airborne contagion: conclusions overgeneralized.
PLoS Med. 2007 May;4(5):e189; author reply e195. [Abstract/Link to Full Text]

Escombe AR, Oeser CC, Gilman RH, Navincopa M, Ticona E, Pan W, Martínez C, Chacaltana J, Rodríguez R, Moore DA, Friedland JS, Evans CA
Natural ventilation for the prevention of airborne contagion.
PLoS Med. 2007 Feb;4(2):e68.
BACKGROUND: Institutional transmission of airborne infections such as tuberculosis (TB) is an important public health problem, especially in resource-limited settings where protective measures such as negative-pressure isolation rooms are difficult to implement. Natural ventilation may offer a low-cost alternative. Our objective was to investigate the rates, determinants, and effects of natural ventilation in health care settings. METHODS AND FINDINGS: The study was carried out in eight hospitals in Lima, Peru; five were hospitals of "old-fashioned" design built pre-1950, and three of "modern" design, built 1970-1990. In these hospitals 70 naturally ventilated clinical rooms where infectious patients are likely to be encountered were studied. These included respiratory isolation rooms, TB wards, respiratory wards, general medical wards, outpatient consulting rooms, waiting rooms, and emergency departments. These rooms were compared with 12 mechanically ventilated negative-pressure respiratory isolation rooms built post-2000. Ventilation was measured using a carbon dioxide tracer gas technique in 368 experiments. Architectural and environmental variables were measured. For each experiment, infection risk was estimated for TB exposure using the Wells-Riley model of airborne infection. We found that opening windows and doors provided median ventilation of 28 air changes/hour (ACH), more than double that of mechanically ventilated negative-pressure rooms ventilated at the 12 ACH recommended for high-risk areas, and 18 times that with windows and doors closed (p < 0.001). Facilities built more than 50 years ago, characterised by large windows and high ceilings, had greater ventilation than modern naturally ventilated rooms (40 versus 17 ACH; p < 0.001). Even within the lowest quartile of wind speeds, natural ventilation exceeded mechanical (p < 0.001). The Wells-Riley airborne infection model predicted that in mechanically ventilated rooms 39% of susceptible individuals would become infected following 24 h of exposure to untreated TB patients of infectiousness characterised in a well-documented outbreak. This infection rate compared with 33% in modern and 11% in pre-1950 naturally ventilated facilities with windows and doors open. CONCLUSIONS: Opening windows and doors maximises natural ventilation so that the risk of airborne contagion is much lower than with costly, maintenance-requiring mechanical ventilation systems. Old-fashioned clinical areas with high ceilings and large windows provide greatest protection. Natural ventilation costs little and is maintenance free, and is particularly suited to limited-resource settings and tropical climates, where the burden of TB and institutional TB transmission is highest. In settings where respiratory isolation is difficult and climate permits, windows and doors should be opened to reduce the risk of airborne contagion. [Abstract/Link to Full Text]

Muula A
Undertaking research in other countries: further considerations.
PLoS Med. 2007 May;4(5):e187; author reply e188. [Abstract/Link to Full Text]

Skene L
Undertaking research in other countries: national ethico-legal barometers and international ethical consensus statements.
PLoS Med. 2007 Feb;4(2):e10.
Is it ethical for scientists to conduct or to benefit from research in another country if that research would be unlawful, or not generally accepted, in their own country? [Abstract/Link to Full Text]

Schackman BR, Neukermans CP, Fontain SN, Nolte C, Joseph P, Pape JW, Fitzgerald DW
Cost-effectiveness of rapid syphilis screening in prenatal HIV testing programs in Haiti.
PLoS Med. 2007 May;4(5):e183.
BACKGROUND: New rapid syphilis tests permit simple and immediate diagnosis and treatment at a single clinic visit. We compared the cost-effectiveness, projected health outcomes, and annual cost of screening pregnant women using a rapid syphilis test as part of scaled-up prenatal testing to prevent mother-to-child HIV transmission in Haiti. METHODS AND FINDINGS: A decision analytic model simulated health outcomes and costs separately for pregnant women in rural and urban areas. We compared syphilis syndromic surveillance (rural standard of care), rapid plasma reagin test with results and treatment at 1-wk follow-up (urban standard of care), and a new rapid test with immediate results and treatment. Test performance data were from a World Health Organization-Special Programme for Research and Training in Tropical Diseases field trial conducted at the GHESKIO Center Groupe Haitien d'Etude du Sarcome de Kaposi et des Infections Opportunistes in Port-au-Prince. Health outcomes were projected using historical data on prenatal syphilis treatment efficacy and included disability-adjusted life years (DALYs) of newborns, congenital syphilis cases, neonatal deaths, and stillbirths. Cost-effectiveness ratios are in US dollars/DALY from a societal perspective; annual costs are in US dollars from a payer perspective. Rapid testing with immediate treatment has a cost-effectiveness ratio of $6.83/DALY in rural settings and $9.95/DALY in urban settings. Results are sensitive to regional syphilis prevalence, rapid test sensitivity, and the return rate for follow-up visits. Integrating rapid syphilis testing into a scaled-up national HIV testing and prenatal care program would prevent 1,125 congenital syphilis cases and 1,223 stillbirths or neonatal deaths annually at a cost of $525,000. CONCLUSIONS: In Haiti, integrating a new rapid syphilis test into prenatal care and HIV testing would prevent congenital syphilis cases and stillbirths, and is cost-effective. A similar approach may be beneficial in other resource-poor countries that are scaling up prenatal HIV testing. [Abstract/Link to Full Text]

Watase K, Gatchel JR, Sun Y, Emamian E, Atkinson R, Richman R, Mizusawa H, Orr HT, Shaw C, Zoghbi HY
Lithium therapy improves neurological function and hippocampal dendritic arborization in a spinocerebellar ataxia type 1 mouse model.
PLoS Med. 2007 May;4(5):e182.
BACKGROUND: Spinocerebellar ataxia type 1 (SCA1) is a dominantly inherited neurodegenerative disorder characterized by progressive motor and cognitive dysfunction. Caused by an expanded polyglutamine tract in ataxin 1 (ATXN1), SCA1 pathogenesis involves a multifactorial process that likely begins with misfolding of ATXN1, which has functional consequences on its interactions, leading to transcriptional dysregulation. Because lithium has been shown to exert neuroprotective effects in a variety of conditions, possibly by affecting gene expression, we tested the efficacy of lithium treatment in a knock-in mouse model of SCA1 (Sca1(154Q/2Q) mice) that replicates many features of the human disease. METHODS AND FINDINGS: Sca1(154Q/2Q) mice and their wild-type littermates were fed either regular chow or chow that contained 0.2% lithium carbonate. Dietary lithium carbonate supplementation resulted in improvement of motor coordination, learning, and memory in Sca1(154Q/2Q) mice. Importantly, motor improvement was seen when treatment was initiated both presymptomatically and after symptom onset. Neuropathologically, lithium treatment attenuated the reduction of dendritic branching in mutant hippocampal pyramidal neurons. We also report that lithium treatment restored the levels of isoprenylcysteine carboxyl methyltransferase (Icmt; alternatively, Pccmt), down-regulation of which is an early marker of mutant ATXN1 toxicity. CONCLUSIONS: The effect of lithium on a marker altered early in the course of SCA1 pathogenesis, coupled with its positive effect on multiple behavioral measures and hippocampal neuropathology in an authentic disease model, make it an excellent candidate treatment for human SCA1 patients. [Abstract/Link to Full Text]

Keen J, Serghides L, Ayi K, Patel SN, Ayisi J, van Eijk A, Steketee R, Udhayakumar V, Kain KC
HIV impairs opsonic phagocytic clearance of pregnancy-associated malaria parasites.
PLoS Med. 2007 May;4(5):e181.
BACKGROUND: Primigravid (PG) women are at risk for pregnancy-associated malaria (PAM). Multigravid (MG) women acquire protection against PAM; however, HIV infection impairs this protective response. Protection against PAM is associated with the production of IgG specific for variant surface antigens (VSA-PAM) expressed by chondroitin sulfate A (CSA)-adhering parasitized erythrocytes (PEs). We hypothesized that VSA-PAM-specific IgG confers protection by promoting opsonic phagocytosis of PAM isolates and that HIV infection impairs this response. METHODS AND FINDINGS: We assessed the ability of VSA-PAM-specific IgG to promote opsonic phagocytosis of CSA-adhering PEs and the impact of HIV infection on this process. Opsonic phagocytosis assays were performed using the CSA-adherent parasite line CS2 and human and murine macrophages. CS2 PEs were opsonized with plasma or purified IgG subclasses from HIV-negative or HIV-infected PG and MG Kenyan women or sympatric men. Levels of IgG subclasses specific for VSA-PAM were compared in HIV-negative and HIV-infected women by flow cytometry. Plasma from HIV-negative MG women, but not PG women or men, promoted the opsonic phagocytosis of CSA-binding PEs (p < 0.001). This function depended on VSA-PAM-specific plasma IgG1 and IgG3. HIV-infected MG women had significantly lower plasma opsonizing activity (median phagocytic index 46 [interquartile range (IQR) 18-195] versus 251 [IQR 93-397], p = 0.006) and levels of VSA-PAM-specific IgG1 (mean fluorescence intensity [MFI] 13 [IQR 11-20] versus 30 [IQR 23-41], p < 0.001) and IgG3 (MFI 17 [IQR 14-23] versus 28 [IQR 23-37], p < 0.001) than their HIV-negative MG counterparts. CONCLUSIONS: Opsonic phagocytosis may represent a novel correlate of protection against PAM. HIV infection may increase the susceptibility of multigravid women to PAM by impairing this clearance mechanism. [Abstract/Link to Full Text]

Fotheringham J, Donati D, Akhyani N, Fogdell-Hahn A, Vortmeyer A, Heiss JD, Williams E, Weinstein S, Bruce DA, Gaillard WD, Sato S, Theodore WH, Jacobson S
Association of human herpesvirus-6B with mesial temporal lobe epilepsy.
PLoS Med. 2007 May;4(5):e180.
BACKGROUND: Human herpesvirus-6 (HHV-6) is a beta-herpesvirus with 90% seroprevalence that infects and establishes latency in the central nervous system. Two HHV-6 variants are known: HHV-6A and HHV-6B. Active infection or reactivation of HHV-6 in the brain is associated with neurological disorders, including epilepsy, encephalitis, and multiple sclerosis. In a preliminary study, we found HHV-6B DNA in resected brain tissue from patients with mesial temporal lobe epilepsy (MTLE) and have localized viral antigen to glial fibrillary acidic protein (GFAP)-positive glia in the same brain sections. We sought, first, to determine the extent of HHV-6 infection in brain material resected from MTLE and non-MTLE patients; and second, to establish in vitro primary astrocyte cultures from freshly resected brain material and determine expression of glutamate transporters. METHODS AND FINDINGS: HHV-6B infection in astrocytes and brain specimens was investigated in resected brain material from MTLE and non-MTLE patients using PCR and immunofluorescence. HHV-6B viral DNA was detected by TaqMan PCR in brain resections from 11 of 16 (69%) additional patients with MTLE and from zero of seven (0%) additional patients without MTLE. All brain regions that tested positive by HHV-6B variant-specific TaqMan PCR were positive for viral DNA by nested PCR. Primary astrocytes were isolated and cultured from seven epilepsy brain resections and astrocyte purity was defined by GFAP reactivity. HHV-6 gp116/54/64 antigen was detected in primary cultured GFAP-positive astrocytes from resected tissue that was HHV-6 DNA positive-the first demonstration of an ex vivo HHV-6-infected astrocyte culture isolated from HHV-6-positive brain material. Previous work has shown that MTLE is related to glutamate transporter dysfunction. We infected astrocyte cultures in vitro with HHV-6 and found a marked decrease in glutamate transporter EAAT-2 expression. CONCLUSIONS: Overall, we have now detected HHV-6B in 15 of 24 patients with mesial temporal sclerosis/MTLE, in contrast to zero of 14 with other syndromes. Our results suggest a potential etiology and pathogenic mechanism for MTLE. [Abstract/Link to Full Text]

Recent Articles in BMC Pharmacology

Chang JY, Liu LZ
Peroxisome proliferator-activated receptor agonists prevent 25-OH-cholesterol induced c-jun activation and cell death.
BMC Pharmacol. 2001;110.
BACKGROUND: Cholesterol oxides, the oxygenated derivatives of cholesterol, have been shown to cause programmed cell death in a variety of cell types. Using N9 microglia, this study was designed to investigate the molecular events induced by cholesterol oxides prior to the execution of programmed cell death. RESULTS: Microglia were very sensitive to 25-OH-cholesterol, such that a 2-day treatment of the cells with 5 microM 25-OH-cholesterol reduced cell viability to 5-10% of controls. There was a dose- and time-dependent increase in c-jun and phospho-c-jun levels in microglia prior to this 25-OH-cholesterol induced cell death. In contrast, 7-beta-OH-cholesterol, which was relatively non-toxic to microglia, did not increase phospho-c-jun levels. Peroxisome proliferator-activated receptors (PPARs) are a group of nuclear receptors that have important roles in atherogenesis. Results from this study indicate that PPAR agonists such as 15d-PGJ2, indomethacin and WY14643 can attenuate cholesterol oxide induced c-jun activation and cell death in microglia. CONCLUSIONS: Peroxisome proliferator-activated receptor agonists may be useful in future development of pharmacological agents against cholesterol oxide induced cytotoxicity. [Abstract/Link to Full Text]

Mayer AM, Hall M, Fay MJ, Lamar P, Pearson C, Prozialeck WC, Lehmann VK, Jacobson PB, Romanic AM, Uz T, Manev H
Effect of a short-term in vitro exposure to the marine toxin domoic acid on viability, tumor necrosis factor-alpha, matrix metalloproteinase-9 and superoxide anion release by rat neonatal microglia.
BMC Pharmacol. 2001;17.
BACKGROUND: The excitatory amino acid domoic acid, a glutamate and kainic acid analog, is the causative agent of amnesic shellfish poisoning in humans. No studies to our knowledge have investigated the potential contribution to short-term neurotoxicity of the brain microglia, a cell type that constitutes circa 10% of the total glial population in the brain. We tested the hypothesis that a short-term in vitro exposure to domoic acid, might lead to the activation of rat neonatal microglia and the concomitant release of the putative neurotoxic mediators tumor necrosis factor-alpha (TNF-alpha), matrix metalloproteinases-2 and-9 (MMP-2 and -9) and superoxide anion (O2-). RESULTS: In vitro, domoic acid [10 microM-1 mM] was significantly neurotoxic to primary cerebellar granule neurons. Although neonatal rat microglia expressed ionotropic glutamate GluR4 receptors, exposure during 6 hours to domoic acid [10 microM-1 mM] had no significant effect on viability. By four hours, LPS (10 ng/mL) stimulated an increase in TNF-alpha mRNA and a 2,233 % increase in TNF-alpha protein In contrast, domoic acid (1 mM) induced a slight rise in TNF-alpha expression and a 53 % increase (p < 0.01) of immunoreactive TNF-alpha protein. Furthermore, though less potent than LPS, a 4-hour treatment with domoic acid (1 mM) yielded a 757% (p < 0.01) increase in MMP-9 release, but had no effect on MMP-2. Finally, while PMA (phorbol 12-myristate 13-acetate) stimulated O2- generation was elevated in 6 hour LPS-primed microglia, a similar pretreatment with domoic acid (1 mM) did not prime O2- release. CONCLUSIONS: To our knowledge this is the first experimental evidence that domoic acid, at in vitro concentrations that are toxic to neuronal cells, can trigger a release of statistically significant amounts of TNF-alpha and MMP-9 by brain microglia. These observations are of considerable pathophysiological significance because domoic acid activates rat microglia several days after in vivo administration. [Abstract/Link to Full Text]

Banes AK, Loberg RD, Brosius FC, Watts SW
Inability of serotonin to activate the c-Jun N-terminal kinase and p38 kinase pathways in rat aortic vascular smooth muscle cells.
BMC Pharmacol. 2001;18.
BACKGROUND: Serotonin (5-HT, 5-hydroxytryptamine) activates the Extracellular Signal-Regulated Kinase (ERK)/ Mitogen-Activated Protein Kinase (MAPK) pathways, in vascular smooth muscle cells. Parallel MAPK pathways, the c-Jun N-terminal Kinase (JNK) and p38 pathway, are activated by stimulators of the ERK/MAPK pathway. We hypothesized that 5-HT would activate the JNK and p38 pathways in rat vascular smooth muscle cells. RESULTS: Results were determined using standard Western analysis and phosphospecific JNK and p38 antibodies. No significant activation by 5-HT (10(-9) - 10(-5) M; 30 min) of the JNK or p38 pathways, as measured by protein phosphorylation, was observed in any of these experiments. These experiments were repeated in the presence of the serine/threonine phosphatase inhibitor okadaic acid (1 uM) and the tyrosine phosphatase inhibitor sodium orthovanadate (1 uM) to maximize any observable signal. Even under these optimized conditions, no activation of the JNK or p38 pathways by 5-HT was observed. Time course experiments (5-HT 10(-5) M; 5 min, 15 min, 30 min and 60 min) showed no significant activation of JNK after incubation with 5-HT at any time point. However, we detected strong activation of JNK p54 and p46 (5- and 7 fold increases in bands p54 and p46, respectively over control levels) by anisomycin (500 ng/ml, 30 min). Similarly, a JNK activity assay failed to reveal activation of JNK by 5-HT, in contrast to the strong stimulation by anisomycin. CONCLUSION: Collectively, these data support the conclusion that 5-HT does not activate the JNK or p38 pathways in rat vascular smooth muscle cells. [Abstract/Link to Full Text]

Morazo P, Fortepiani LA, Clara Ortíz M, Atucha NM, García-Estań J
Omapatrilat normalizes renal function curve in spontaneously hypertensive rats.
BMC Pharmacol. 2001;15.
BACKGROUND: The present study was designed to analyze the chronic renal response to omapatrilat, a new vasopeptidase inhibitor, in spontaneously hypertensive rats (SHR). To that end, the renal and blood pressure response to a 4-day salt loading protocol was analyzed and the respective chronic renal curves constructed. RESULTS: In non treated animals, and under normal sodium intake (around 2 mEq/day), mean arterial pressure (MAP), was significantly higher in the SHR as compared with the controls (WKY). After increasing salt intake (8 times normal), MAP did not change significantly in any group and the animals reached a normal sodium balance in four days. In a second group of animals, omapatrilat was given orally for 15 days at the dose of 40 mg/kg/day in the drinking water. In these omapatrilat-treated animals, and under normal sodium intake, MAP was significantly lower in both groups, although the antihypertensive effect was much greater in the SHR, so that the MAP of the SHR group was completely normalized and similar to the WKY-treated group. The subsequent elevation of sodium intake did not significantly elevate MAP in any group and the animals could manage the sodium excess as well as the non treated groups. CONCLUSIONS: These results indicate that chronic treatment with omapatrilat normalizes blood pressure in SHR without affecting adversely the renal ability to eliminate a sodium load. Chronic treatment with omapatrilat resets the chronic pressure natriuresis relationship of the SHR to a normal level, thus without altering the normal salt-independence of this arterial hypertension model. [Abstract/Link to Full Text]

Vera PL, Miranda-Sousa A, Nadelhaft I
Effects of two atypical neuroleptics, olanzapine and risperidone, on the function of the urinary bladder and the external urethral sphincter in anesthetized rats.
BMC Pharmacol. 2001;14.
BACKGROUND: A previous report showed that the atypical neuroleptic clozapine resulted in marked changes in urodynamic parameters and greatly inhibited the activity of the external urethral sphincter in anesthetized rats. Such findings may help explain the high incidence of urinary disturbances reported during clozapine therapy. In an effort to extend our observations to other atypical neuroleptic agents, the present study investigated the effects of two newer atypical antipsychotics, olanzapine and risperidone, on the bladder and external urethral sphincter during cystometry in anesthetized rats. RESULTS: At a dose of 0.1 mg/kg (i.v.), olanzapine decreased the micturition volume and increased the residual volume. In addition, olanzapine decreased the expulsion time and the amplitude of the high frequency oscillations observed during the expulsion phase. Larger doses (1 mg/kg) had a greater effect. Olanzapine also reduced the activity recorded from the external urethral sphincter, and the bursting observed during the expulsion phase was abolished by 1.0 mg/kg. Risperidone had similar effects although the maximal effects were smaller than those observed with olanzapine. The amplitude of bladder contractions elicited by electrical stimulation of the pelvic nerve was reduced by olanzapine but not risperidone suggesting a possible anti-muscarinic peripheral effect of olanzapine. CONCLUSIONS: Olanzapine and risperidone significantly altered several voiding parameters and decreased the activity of the external urethral sphincter in the anesthetized rat. We propose that these effects are due to the central action of these drugs and not to peripheral effects. These findings may explain some of the clinical reports of urinary incontinence with risperidone and may predict similar occurrences with olanzapine therapy. [Abstract/Link to Full Text]

de Miranda FG, Vilar JC, Alves IA, Cavalcanti SC, Antoniolli AR
Antinociceptive and antiedematogenic properties and acute toxicity of Tabebuia avellanedae Lor. ex Griseb. inner bark aqueous extract.
BMC Pharmacol. 2001;16.
BACKGROUND: Tabebuia avellanedae is a tree from the Bignoniaceae family. Commonly know as "pau d'arco" in Brazil, its inner bark is used as analgesic, anti-inflammatory, antineoplasic and diuretic at the Brazilian northeast. A validation of the plant usage has not been previously performed. RESULTS: Antinociceptive and antiedematogenic effects of Tabebuia avellanedae Lor. ex Griseb. inner bark were measured by nociceptive experimental models in mice. A rat paw edema test induced by carrageenan (1%) was also performed in rats to access the plant's antiedematogenic effect. The inner bark aqueous extract, administered via oral in three different concentration, namely 100, 200 and 400 mg/Kg, reduced the nociception produced by acetic acid (0.6% in water, i.p.) by 49.9%, 63.7% and 43.8%, respectively. The aqueous extract (200 and 400 mg/Kg, p.o.) reduced formalin (1%) effects only at the second phase of the experiment by 49.3% and 53.7%, respectively. Naloxone (5 mg/Kg, i.p.) was not able to revert the extract effect, however caffeine (10 mg/Kg, i.p.) reverted its effect by 19.8% at the second phase of the formalin test. The aqueous extract (200 mg/Kg, p.o.) inhibited edema by 12.9% when we used the rat paw edema model. The acute toxicity was low in mice. CONCLUSION: The T. avellanedae inner bark aqueous extract presented antinociceptive and antiedematogenic activities at the used models, with a possible antinociceptive effect associated to the adenosine system. [Abstract/Link to Full Text]

Suga S, Wu J, Ogawa Y, Takeo T, Kanno T, Wakui M
Phorbol ester impairs electrical excitation of rat pancreatic beta-cells through PKC-independent activation of KATP channels.
BMC Pharmacol. 2001;13.
BACKGROUND: Phorbol 12-myristate 13-acetate (PMA) is often used as an activating phorbol ester of protein kinase C (PKC) to investigate the roles of the kinase in cellular functions. Accumulating lines of evidence indicate that in addition to activating PKC, PMA also produces some regulatory effects in a PKC-independent manner. In this study, we investigated the non-PKC effects of PMA on electrical excitability of rat pancreatic beta-cells by using patch-clamp techniques. RESULTS: In current-clamp recording, PMA (80 nM) reversibly inhibited 15 mM glucose-induced action potential spikes superimposed on a slow membrane depolarization and this inhibition can not be prevented by pre-treatment of the cell with a specific PKC inhibitor, bisindolylmaleimide (BIM, 1 microM). In the presence of a subthreshold concentration (5.5 mM) of glucose, PMA hyperpolarized beta-cells in a concentration-dependent manner (0.8-240 nM), even in the presence of BIM. Based on cell-attached single channel recordings, PMA increased ATP-sensitive K+ channel (KATP) activity. Based on inside-out patch-clamp recordings, PMA had little effect on KATP activity if no ATP was in the bath, while PMA restored KATP activity that was suppressed by 10 microM ATP in the bath. In voltage-clamp recording, PMA enhanced tolbutamide-sensitive membrane currents elicited by repetitive ramp pulses from -90 to -50 mV in a concentration-dependent manner, and this potentiation could not be prevented by pre-treatment of cell with BIM. 4alpha-phorbol 12,13-didecanoate (4alpha-PDD), a non-PKC-activating phorbol ester, mimicked the effect of PMA on both current-clamp and voltage-clamp recording configurations. With either 5.5 or 16.6 mM glucose in the extracellular solution, PMA (80 nM) increased insulin secretion from rat islets. However, in islets pretreated with BIM (1 microM), PMA did not increase, but rather reduced insulin secretion. CONCLUSION: In rat pancreatic beta-cells, PMA modulates insulin secretion through a mixed mechanism: increases insulin secretion by activation of PKC, and meanwhile decrease insulin secretion by impairing beta-cell excitability in a PKC-independent manner. The enhancement of KATP activity by reducing sensitivity of KATP to ATP seems to underlie the PMA-induced impairment of beta-cells electrical excitation in response to glucose stimulation. [Abstract/Link to Full Text]

Briner W
The effect of GABA receptor ligands in experimental spina bifida occulta.
BMC Pharmacol. 2001;12.
BACKGROUND: The pathophysiology behind spina bifida and other neural tube defects (NTDs) is unclear. Folic acid is one variable, but other factors remain. Studies suggest that substances active at the GABA receptor may produce NTDs. To test this hypothesis pregnant rats were exposed to either the GABA a agonist muscimol (1, 2 or 4 mg/kg), the GABA a antagonist bicuculline (.5, 1, or 2 mg/kg), the GABA b agonist baclofen (15, 30, 60 mg/kg), or the GABA b antagonist hydroxysaclofen (1, 3, or 5 mg/kg) during neural tube formation. Normal saline was used as a control and valproic acid (600 mg/kg) as a positive control. The embryos were analyzed for the presence of a spina bifida like NTD. RESULTS: After drug administration the pregnancies were allowed to proceed to the 21st day of gestation. Then embryos were removed and skeletons staining and cleared. Vertebral arch closure was measured. Results indicate that the GABAa receptor agonist muscimol, the GABAa receptor antagonist bicuculline, and the GABAb agonist baclofen produced NTDs characterized by widening of the vertebral arch. Oppositely the GABAb antagonist hydroxysaclofen produced narrowing of the vertebral arches. CONCLUSIONS: The findings indicate that GABA a or b ligands are capable of altering neural formation. GABA may play a greater than appreciated role in neural tube formation and may be important in NTDs. The narrowing of the vertebral arch produced by the GABA b antagonist hydroxysalcofen suggests that GABA b receptor may play an undefined role in neural tube closure that differs from the GABA a receptor. [Abstract/Link to Full Text]

Kamau SW, Nunez R, Grimm F
Flow cytometry analysis of the effect of allopurinol and the dinitroaniline compound (Chloralin) on the viability and proliferation of Leishmania infantum promastigotes.
BMC Pharmacol. 2001;11.
BACKGROUND: Leishmaniasis is a major parasitic disease in the tropical regions. However, Leishmania infantum has recently emerged as a very important cause of opportunistic infections for individuals positive for human immunodeficiency virus (HIV). However, there is a lack of in vitro tests for assessing the effect of anti-parasitic drugs on the viability and proliferation of Leishmania infantum. The aim of this study is to assess the efficacy of anti-parasitic drugs like allopurinol and Chloralin on the viability and proliferation of L. infantum promastigotes by utilizing two complementary flow cytometric approaches after exposure of the promastigotes to various concentrations of the drugs. RESULTS: The density of the cultures in the presence and absence of allopurinol was determined by haemocytometer enumeration. The two flow cytometric approaches used to monitor the drug effect were: (i) a quantitative method to measure cell division using 5-,6-carboxyfluorescein diacetate succinimidyl ester (CFSE) staining and (ii) evaluation of cell viability by dual-staining with the membrane-permeable nuclear stain, SBRY-14 and propidium iodide. It was found that concentrations of allopurinol above 50 microg/ml yielded a clear decrease in the proliferation rate of the promastigotes. However, the viability results showed that about 46.8% of the promastigotes incubated in the presence of 800 microg/ml of allopurinol were still alive after 96 hours. In sharp contrast, more than 90% of promastigotes treated with Chloralin 10 microM (2.7 microg/ml) were dead after 48 hours of treatment. These flow cytometric findings suggest that allopurinol has a leishmaniostatic effect while the dinitroaniline compound (Chloralin) has a leishmaniocidal effect against promastigotes. CONCLUSIONS: The flow cytometric data on proliferation and viability were consistent with results obtained from haemocytometer counts and allowed us to develop a model for assessing in vitro the effects of medicaments like allopurinol and chloralin on L. infantum promastigotes on a cellular level. [Abstract/Link to Full Text]

Recent Articles in BMC Clinical Pharmacology

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Recent Articles in BMC Medicine

Lalloo AK, Luo FR, Guo A, Paranjpe PV, Lee SH, Vyas V, Rubin E, Sinko PJ
Membrane transport of camptothecin: facilitation by human P-glycoprotein (ABCB1) and multidrug resistance protein 2 (ABCC2).
BMC Med. 2004 May 4;216.
BACKGROUND: The purpose of the present study was to continue the investigation of the membrane transport mechanisms of 20-(S)-camptothecin (CPT) in order to understand the possible role of membrane transporters on its oral bioavailability and disposition. METHODS: The intestinal transport kinetics of CPT were characterized using Caco-2 cells, MDCKII wild-type cells and MDCKII cells transfected with human P-glycoprotein (PGP) (ABCB1) or human multidrug resistance protein 2 (MRP2) (ABCC2). The effects of drug concentration, inhibitors and temperature on CPT directional permeability were determined. RESULTS: The absorptive (apical to basolateral) and secretory (basolateral to apical) permeabilities of CPT were found to be saturable. Reduced secretory CPT permeabilities with decreasing temperatures suggests the involvement of an active, transporter-mediated secretory pathway. In the presence of etoposide, the CPT secretory permeability decreased 25.6%. However, inhibition was greater in the presence of PGP and of the breast cancer resistant protein inhibitor, GF120918 (52.5%). The involvement of additional secretory transporters was suggested since the basolateral to apical permeability of CPT was not further reduced in the presence of increasing concentrations of GF120918. To investigate the involvement of specific apically-located secretory membrane transporters, CPT transport studies were conducted using MDCKII/PGP cells and MDCKII/MRP2 cells. CPT carrier-mediated permeability was approximately twofold greater in MDCKII/PGP cells and MDCKII/MRP2 cells than in MDCKII/wild-type cells, while the apparent Km values were comparable in all three cell lines. The efflux ratio of CPT in MDCKII/PGP in the presence of 0.2 microM GF120918 was not completely reversed (3.36 to 1.49). However, the decrease in the efflux ratio of CPT in MDCKII/MRP2 cells (2.31 to 1.03) suggests that CPT efflux was completely inhibited by MK571, a potent inhibitor of the Multidrug Resistance Protein transporter family. CONCLUSIONS: The current results provide evidence that PGP and MRP2 mediate the secretory transport of CPT in vitro. However, the involvement of other transporters cannot be ruled out based on these studies. Since these transporters are expressed in the intestine, liver and kidney variations in their expression levels and/or regulation may be responsible for the erratic oral absorption and biliary excretion of CPT observed in human subjects. [Abstract/Link to Full Text]

McGrath J, Saha S, Welham J, El Saadi O, MacCauley C, Chant D
A systematic review of the incidence of schizophrenia: the distribution of rates and the influence of sex, urbanicity, migrant status and methodology.
BMC Med. 2004 Apr 28;213.
BACKGROUND: Understanding variations in the incidence of schizophrenia is a crucial step in unravelling the aetiology of this group of disorders. The aims of this review are to systematically identify studies related to the incidence of schizophrenia, to describe the key features of these studies, and to explore the distribution of rates derived from these studies. METHODS: Studies with original data related to the incidence of schizophrenia (published 1965-2001) were identified via searching electronic databases, reviewing citations and writing to authors. These studies were divided into core studies, migrant studies, cohort studies and studies based on Other Special Groups. Between- and within-study filters were applied in order to identify discrete rates. Cumulative plots of these rates were made and these distributions were compared when the underlying rates were sorted according to sex, urbanicity, migrant status and various methodological features. RESULTS: We identified 100 core studies, 24 migrant studies, 23 cohort studies and 14 studies based on Other Special Groups. These studies, which were drawn from 33 countries, generated a total of 1,458 rates. Based on discrete core data for persons (55 studies and 170 rates), the distribution of rates was asymmetric and had a median value (10%-90% quantile) of 15.2 (7.7-43.0) per 100,000. The distribution of rates was significantly higher in males compared to females; the male/female rate ratio median (10%-90% quantile) was 1.40 (0.9-2.4). Those studies conducted in urban versus mixed urban-rural catchment areas generated significantly higher rate distributions. The distribution of rates in migrants was significantly higher compared to native-born; the migrant/native-born rate ratio median (10%-90% quantile) was 4.6 (1.0-12.8). Apart from the finding that older studies reported higher rates, other study features were not associated with significantly different rate distributions (e.g. overall quality, methods related to case finding, diagnostic confirmation and criteria, the use of age-standardization and age range). CONCLUSIONS: There is a wealth of data available on the incidence of schizophrenia. The width and skew of the rate distribution, and the significant impact of sex, urbanicity and migrant status on these distributions, indicate substantial variations in the incidence of schizophrenia. [Abstract/Link to Full Text]

Goodwin JL, Kaemingk KL, Fregosi RF, Rosen GM, Morgan WJ, Smith T, Quan SF
Parasomnias and sleep disordered breathing in Caucasian and Hispanic children - the Tucson children's assessment of sleep apnea study.
BMC Med. 2004 Apr 28;214.
BACKGROUND: Recent studies in children have demonstrated that frequent occurrence of parasomnias is related to increased sleep disruption, mental disorders, physical harm, sleep disordered breathing, and parental duress. Although there have been several cross-sectional and clinical studies of parasomnias in children, there have been no large, population-based studies using full polysomnography to examine the association between parasomnias and sleep disordered breathing. The Tucson Children's Assessment of Sleep Apnea study is a community-based cohort study designed to investigate the prevalence and correlates of objectively measured sleep disordered breathing (SDB) in pre-adolescent children six to 11 years of age. This paper characterizes the relationships between parasomnias and SDB with its associated symptoms in these children. METHODS: Parents completed questionnaires pertaining to their child's sleep habits. Children had various physiological measurements completed and then were connected to the Compumedics PS-2 sleep recording system for full, unattended polysomnography in the home. A total of 480 unattended home polysomnograms were completed on a sample that was 50% female, 42.3% Hispanic, and 52.9% between the ages of six and eight years. RESULTS: Children with a Respiratory Disturbance Index of one or greater were more likely to have sleep walking (7.0% versus 2.5%, p < 0.02), sleep talking (18.3% versus 9.0%, p < 0.006), and enuresis (11.3% versus 6.3%, p < 0.08) than children with an Respiratory Disturbance Index of less than one. A higher prevalence of other sleep disturbances as well as learning problems was observed in children with parasomnia. Those with parasomnias associated with arousal were observed to have increased number of stage shifts. Small alterations in sleep architecture were found in those with enuresis. CONCLUSIONS: In this population-based cohort study, pre-adolescent school-aged children with SDB experienced more parasomnias than those without SDB. Parasomnias were associated with a higher prevalence of other sleep disturbances and learning problems. Clinical evaluation of children with parasomnias should include consideration of SDB. [Abstract/Link to Full Text]

Ather MH, Faruqui N, Akhtar S, Sulaiman MN
Is an excretory urogram mandatory in patients with small to medium-sized renal and ureteric stones treated by extra corporeal shock wave lithotripsy?
BMC Med. 2004 Apr 28;215.
BACKGROUND: An intravenous urogram (IVU) has traditionally been considered mandatory before treating renal and ureteric stones by extracorporeal shock wave lithotripsy (ESWL). This study was designed to see whether there is a difference in complications and the need for ancillary procedures in patients managed by ESWL for renal and ureteric calculi, according to preoperative imaging technique. METHODS: This retrospective study compared 133 patients undergoing ESWL from January 2001 to July 2002. Patients were divided into three groups according to the preoperative imaging technique used: i) IVU; ii) non-contrast enhanced helical computed tomography (UHCT); and iii) ultrasound (US) + X-ray kidney, ureter and bladder (KUB). The groups were matched in terms of age and gender, as well as location, side and size of stones. RESULTS: There was no statistically significantly difference for number of ESWL sessions, number of shock waves and use of ancillary procedures between the three groups. The stone-free rate was 98% for the IVU and UHCT groups, and 97% for the US + X-ray KUB group. CONCLUSIONS: The complication rate and need for ancillary procedures was comparable across the three groups. Patients imaged by UHCT or US + X-ray KUB prior to ESWL for uncomplicated renal and ureteric stones do not require IVU. [Abstract/Link to Full Text]

Herndon TO, Gonzalez S, Gowrishankar TR, Anderson RR, Weaver JC
Transdermal microconduits by microscission for drug delivery and sample acquisition.
BMC Med. 2004 Apr 19;212.
BACKGROUND: Painless, rapid, controlled, minimally invasive molecular transport across human skin for drug delivery and analyte acquisition is of widespread interest. Creation of microconduits through the stratum corneum and epidermis is achieved by stochastic scissioning events localized to typically 250 microm diameter areas of human skin in vivo. METHODS: Microscissioning is achieved by a limited flux of accelerated gas: 25 microm inert particles passing through the aperture in a mask held against the stratum corneum. The particles scize (cut) tissue, which is removed by the gas flow with the sensation of a gentle stream of air against the skin. The resulting microconduit is fully open and may be between 50 and 200 microm deep. RESULTS: In vivo adult human tests show that microconduits reduce the electrical impedance between two ECG electrodes from approximately 4,000 Omega to 500 Omega. Drug delivery has been demonstrated in vivo by applying lidocaine to a microconduit from a cotton swab. Sharp point probing demonstrated full anaesthesia around the site within three minutes. Topical application without the microconduit required approximately 1.5 hours. Approximately 180 microm deep microconduits in vivo yielded blood sample volumes of several microl, with a faint pricking sensation as blood enters tissue. Blood glucose measurements were taken with two commercial monitoring systems. Microconduits are invisible to the unaided eye, developing a slight erythematous macule that disappears over days. CONCLUSION: Microscissioned microconduits may provide a minimally invasive basis for delivery of any size molecule, and for extraction of interstitial fluid and blood samples. Such microconduits reduce through-skin electrical impedance, have controllable diameter and depth, are fully open and, after healing, no foreign bodies were visible using through-skin confocal microscopy. In subjects to date, microscissioning is painless and rapid. [Abstract/Link to Full Text]

Bellemare S, Hartling L, Wiebe N, Russell K, Craig WR, McConnell D, Klassen TP
Oral rehydration versus intravenous therapy for treating dehydration due to gastroenteritis in children: a meta-analysis of randomised controlled trials.
BMC Med. 2004 Apr 15;211.
BACKGROUND: Despite treatment recommendations from various organizations, oral rehydration therapy (ORT) continues to be underused, particularly by physicians in high-income countries. We conducted a systematic review of randomised controlled trials (RCTs) to compare ORT and intravenous therapy (IVT) for the treatment of dehydration secondary to acute gastroenteritis in children. METHODS: RCTs were identified through MEDLINE, EMBASE, CENTRAL, authors and references of included trials, pharmaceutical companies, and relevant organizations. Screening and inclusion were performed independently by two reviewers in order to identify randomised or quasi-randomised controlled trials comparing ORT and IVT in children with acute diarrhea and dehydration. Two reviewers independently assessed study quality using the Jadad scale and allocation concealment. Data were extracted by one reviewer and checked by a second. The primary outcome measure was failure of rehydration. We analyzed data using standard meta-analytic techniques. RESULTS: The quality of the 14 included trials ranged from 0 to 3 (Jadad score); allocation concealment was unclear in all but one study. Using a random effects model, there was no significant difference in treatment failures (risk difference [RD] 3%; 95% confidence intervals [CI]: 0, 6). The Mantel-Haenzsel fixed effects model gave a significant difference between treatment groups (RD 4%; 95% CI: 2, 5) favoring IVT. Based on the four studies that reported deaths, there were six in the IVT groups and two in ORT. There were no significant differences in total fluid intake at six and 24 hours, weight gain, duration of diarrhea, or hypo/hypernatremia. Length of stay was significantly shorter for the ORT group (weighted mean difference [WMD] -1.2 days; 95% CI: -2.4,-0.02). Phlebitis occurred significantly more often with IVT (number needed to treat [NNT] 33; 95% CI: 25,100); paralytic ileus occurred more often with ORT (NNT 33; 95% CI: 20,100). These results may not be generalizable to children with persistent vomiting. CONCLUSION: There were no clinically important differences between ORT and IVT in terms of efficacy and safety. For every 25 children (95% CI: 20, 50) treated with ORT, one would fail and require IVT. The results support existing practice guidelines recommending ORT as the first course of treatment in appropriate children with dehydration secondary to gastroenteritis. [Abstract/Link to Full Text]

Sinha R, Sharma N, Vajpayee RB
Visual outcome of cataract surgery with pupillary sphincterotomy in eyes with coexisting corneal opacity.
BMC Med. 2004 Apr 7;210.
BACKGROUND: To evaluate the visual outcome following cataract surgery with pupillary sphincterotomy in eyes with coexisting corneal opacity. METHODS: Patients with leucomatous corneal opacity with significant cataract were enrolled for the study. The uncorrected visual acuity and best-corrected visual acuity (BCVA) were recorded and the anterior segment was thoroughly evaluated by a slit lamp biomicroscope before the surgery. Only those patients who had some amount of clear peripheral cornea were selected. Posterior segment pathology was ruled out by indirect ophthalmoscopy after pupillary dilatation, if possible, or by B-scan ultrasonography. Conventional extracapsular cataract extraction with pupillary sphincterotomy was performed and an intraocular lens was implanted. Postoperatively, the eyes were evaluated on day 1, and 1 week and 6 weeks following surgery for similar parameters. RESULTS: Fourteen eyes of 14 patients were included in the study, of which 13 (92.85%) patients were male. The mean age of the patients was 47.85 +/- 7.37 years. All the eyes had a dense central leucomatous corneal opacity. Twelve (85.71%) eyes had two or more quadrants of deep vascularisation. Sphincterotomy was performed mostly (71.42%) in the nasal or inferonasal quadrant. The intraocular lens was implanted in 13 (92.85%) eyes, and one (7.1%) eye was left aphakic due to the occurrence of a large posterior capsular tear. Preoperatively, all eyes had BCVA < 6/60. At 6 weeks after surgery, all eyes had BCVA >or= 6/60 and four (28.57%) eyes had BCVA >or= 6/18. The mean BCVA preoperatively in these eyes was 0.015 +/- 0.009, which changed to 0.249 +/- 0.102 at 6 weeks following surgery. CONCLUSIONS: Extracapsular cataract extraction and intraocular lens implantation with pupillary sphincterotomy provides ambulatory and useful vision to patients of cataract with coexisting central leucomatous corneal opacity. [Abstract/Link to Full Text]

Sugino T, Yamaguchi T, Ogura G, Saito A, Hashimoto T, Hoshi N, Yoshida S, Goodison S, Suzuki T
Morphological evidence for an invasion-independent metastasis pathway exists in multiple human cancers.
BMC Med. 2004 Apr 5;29.
BACKGROUND: We have previously described an alternative invasion-independent pathway of cancer metastasis in a murine mammary tumor model. This pathway is initiated by intravasation of tumor nests enveloped by endothelial cells of sinusoidal vasculature within the tumor. In this study, we examined whether evidence for the invasion-independent pathway of metastasis is present in human cancers. METHODS: Archival specimens of 10 common types of human cancers were examined for the presence of sinusoidal vasculature enveloping tumor nests and subsequently generated endothelial-covered tumor emboli in efferent veins. RESULTS: A percentage of tumor emboli in all cancers was found to be enveloped by endothelial cells, but these structures were particularly prevalent in renal cell carcinomas, hepatocellular carcinomas and follicular thyroid carcinomas. A common feature of the vasculature in these tumors was the presence of dilated sinusoid-like structures surrounding tumor nests. A high mean vascular area within tumors, an indication of sinusoidal vascular development, was significantly related to the presence of endothelial-covered tumor emboli. CONCLUSIONS: These results suggest that an invasion-independent metastatic pathway is possible in a wide variety of human cancers. Further investigation of this phenomenon may present new therapeutic strategies for the amelioration of cancer metastasis. [Abstract/Link to Full Text]

Englund G, Hallberg P, Artursson P, Michaëlsson K, Melhus H
Association between the number of coadministered P-glycoprotein inhibitors and serum digoxin levels in patients on therapeutic drug monitoring.
BMC Med. 2004 Apr 2;28.
BACKGROUND: The ABC transporter P-glycoprotein (P-gp) is recognized as a site for drug-drug interactions and provides a mechanistic explanation for clinically relevant pharmacokinetic interactions with digoxin. The question of whether several P-gp inhibitors may have additive effects has not yet been addressed. METHODS: We evaluated the effects on serum concentrations of digoxin (S-digoxin) in 618 patients undergoing therapeutic drug monitoring. P-gp inhibitors were classified as Class I, with a known effect on digoxin kinetics, or Class II, showing inhibition in vitro but no documented effect on digoxin kinetics in humans. Mean S-digoxin values were compared between groups of patients with different numbers of coadministered P-gp inhibitors by a univariate and a multivariate model, including the potential covariates age, sex, digoxin dose and total number of prescribed drugs. RESULTS: A large proportion (47%) of the digoxin patients undergoing therapeutic drug monitoring had one or more P-gp inhibitor prescribed. In both univariate and multivariate analysis, S-digoxin increased in a stepwise fashion according to the number of coadministered P-gp inhibitors (all P values < 0.01 compared with no P-gp inhibitor). In multivariate analysis, S-digoxin levels were 1.26 +/- 0.04, 1.51 +/- 0.05, 1.59 +/- 0.08 and 2.00 +/- 0.25 nmol/L for zero, one, two and three P-gp inhibitors, respectively. The results were even more pronounced when we analyzed only Class I P-gp inhibitors (1.65 +/- 0.07 for one and 1.83 +/- 0.07 nmol/L for two). CONCLUSIONS: Polypharmacy may lead to multiple drug-drug interactions at the same site, in this case P-gp. The S-digoxin levels increased in a stepwise fashion with an increasing number of coadministered P-gp inhibitors in patients taking P-gp inhibitors and digoxin concomitantly. As coadministration of digoxin and P-gp inhibitors is common, it is important to increase awareness about P-gp interactions among prescribing clinicians. [Abstract/Link to Full Text]

Huber AM, King J, McLaine P, Klassen T, Pothos M
A randomized, placebo-controlled trial of prednisone in early Henoch Schönlein Purpura [ISRCTN85109383].
BMC Med. 2004 Apr 2;27.
BACKGROUND: Henoch Schönlein Purpura (HSP) is the most common systemic vasculitis of childhood. There is considerable controversy over whether children with HSP should be treated with corticosteroids. The goal of this study was to investigate whether early corticosteroid administration could reduce the rate of renal or gastrointestinal complications in children with HSP. METHODS: Forty children with HSP, seen in the emergency room of a tertiary-care, paediatric centre, entered a randomized, double-blind, placebo controlled study. The treatment group (n = 21) received oral prednisone, 2 mg/kg/day for one week, with weaning over a second week, while the placebo group (n = 19) received an identical appearing placebo. Co-primary outcomes were the rate of renal involvement at one year and the rate of acute gastrointestinal complications. Co-primary outcomes were analysed using Fisher's Exact test. RESULTS: At one year, there was no difference in the rate of renal involvement (3/21 prednisone group vs. 2/19 placebo group, P = 1.0). There was also no statistically significant difference in the rate of acute gastrointestinal complications (2/21 prednisone group vs. 3/19 placebo group, P = 0.7). Two children in the placebo group did experience intussusceptions compared with none in the prednisone group (P = 0.2). CONCLUSIONS: Early prednisone therapy in HSP does not appear to reduce the risk of renal involvement at one year, or the risk of acute gastrointestinal complications. There may be a reduced risk of intussusception. The routine, early use of prednisone in uncomplicated HSP cannot be recommended at this time. [Abstract/Link to Full Text]

Sokal D, Irsula B, Hays M, Chen-Mok M, Barone MA
Vasectomy by ligation and excision, with or without fascial interposition: a randomized controlled trial [ISRCTN77781689].
BMC Med. 2004 Mar 31;26.
BACKGROUND: Randomized controlled trials comparing different vasectomy occlusion techniques are lacking. Thus, this multicenter randomized trial was conducted to compare the probability of the success of ligation and excision vasectomy with, versus without, fascial interposition (i.e. placing a layer of the vas sheath between two cut ends of the vas). METHODS: The trial was conducted between December 1999 and June 2002 with a single planned interim analysis. Men requesting vasectomies at eight outpatient clinics in seven countries in North America, Latin America, and Asia were included in the study. The men were randomized to receive vasectomy with versus without fascial interposition. All surgeons performed the vasectomies using the no-scalpel approach to the vas. Participants had a semen analysis two weeks after vasectomy and then every four weeks up to 34 weeks. The primary outcome measure was time to azoospermia. Additional outcome measures were time to severe oligozoospermia (<100 000 sperm/mL) and vasectomy failure based on semen analyses. RESULTS: We halted recruitment after the planned interim analysis, when 841 men had been enrolled. Fascial interposition decreased time to azoospermia (hazard ratio [HR], 1.35; P < 0.0001) and time to severe oligozoospermia (HR, 1.32; P < 0.0001) and reduced failures based on semen analysis by about half, from 12.7% (95% confidence interval [CI], 9.7 to 16.3) to 5.9% (95% CI, 3.8 to 8.6) (P < 0.0001). Older men benefited less from fascial interposition than younger men in terms of the speed of achieving azoospermia. However, the number of vasectomy failures was reduced to a similar degree in all age groups. Slightly more adverse events occurred in the fascial interposition group, but the difference was not significant. These failure rates may appear high to practitioners in countries such as the USA, but they are similar to results from other careful studies of ligation and excision techniques. CONCLUSION: Fascial interposition significantly improves vasectomy success when ligation and excision is the method of vas occlusion. A limitation of this study is that the correlation between postvasectomy sperm concentrations and risk of pregnancy is not well quantified. [Abstract/Link to Full Text]

Lea RA, Ovcaric M, Sundholm J, MacMillan J, Griffiths LR
The methylenetetrahydrofolate reductase gene variant C677T influences susceptibility to migraine with aura.
BMC Med. 2004 Feb 12;23.
BACKGROUND: The C677T variant in the methylenetetrahydrofolate reductase (MTHFR) gene is associated with increased levels of circulating homocysteine and is a mild risk factor for vascular disease. Migraine, with and without aura (MA and MO), is a prevalent and complex neurovascular disorder that may also be affected by genetically influenced hyperhomocysteinaemia. To determine whether the C677T variant in the MTHFR gene is associated with migraine susceptibility we utilised unrelated and family-based case-control study designs. METHODS: A total of 652 Caucasian migraine cases were investigated in this study. The MTHFR C677T variant was genotyped in 270 unrelated migraine cases and 270 controls as well as 382 affected subjects from 92 multiplex pedigrees. RESULTS: In the unrelated case-control sample we observed an over-representation of the 677T allele in migraine patients compared to controls, specifically for the MA subtype (40% vs. 33%) (chi2 = 5.70, P = 0.017). The Armitage test for trend indicated a significant dosage effect of the risk allele (T) for MA (chi2 = 5.72, P = 0.017). This linear trend was also present in the independent family-based sample (chi2 = 4.25, Padjusted = 0.039). Overall, our results indicate that the T/T genotype confers a modest, yet significant, increase in risk for the MA subtype (odds ratio: 2.0 - 2.5). No increased risk for the MO subtype was observed (P > 0.05). CONCLUSIONS: In Caucasians, the C677T variant in the MTHFR gene influences susceptibility to MA, but not MO. Investigation into the enzyme activity of MTHFR and the role of homocysteine in the pathophysiology of migraine is warranted. [Abstract/Link to Full Text]

Klett EL, Lu K, Kosters A, Vink E, Lee MH, Altenburg M, Shefer S, Batta AK, Yu H, Chen J, Klein R, Looije N, Oude-Elferink R, Groen AK, Maeda N, Salen G, Patel SB
A mouse model of sitosterolemia: absence of Abcg8/sterolin-2 results in failure to secrete biliary cholesterol.
BMC Med. 2004 Mar 24;25.
BACKGROUND: Mutations in either of two genes comprising the STSL locus, ATP-binding cassette (ABC)-transporters ABCG5 (encoding sterolin-1) and ABCG8 (encoding sterolin-2), result in sitosterolemia, a rare autosomal recessive disorder of sterol trafficking characterized by increased plasma plant sterol levels. Based upon the genetics of sitosterolemia, ABCG5/sterolin-1 and ABCG8/sterolin-2 are hypothesized to function as obligate heterodimers. No phenotypic difference has yet been described in humans with complete defects in either ABCG5 or ABCG8. These proteins, based upon the defects in humans, are responsible for regulating dietary sterol entry and biliary sterol secretion. METHODS: In order to mimic the human disease, we created, by a targeted disruption, a mouse model of sitosterolemia resulting in Abcg8/sterolin-2 deficiency alone. Homozygous knockout mice are viable and exhibit sitosterolemia. RESULTS: Mice deficient in Abcg8 have significantly increased plasma and tissue plant sterol levels (sitosterol and campesterol) consistent with sitosterolemia. Interestingly, Abcg5/sterolin-1 was expressed in both liver and intestine in Abcg8/sterolin-2 deficient mice and continued to show an apical expression. Remarkably, Abcg8 deficient mice had an impaired ability to secrete cholesterol into bile, but still maintained the ability to secrete sitosterol. We also report an intermediate phenotype in the heterozygous Abcg8+/- mice that are not sitosterolemic, but have a decreased level of biliary sterol secretion relative to wild-type mice. CONCLUSION: These data indicate that Abcg8/sterolin-2 is necessary for biliary sterol secretion and that loss of Abcg8/sterolin-2 has a more profound effect upon biliary cholesterol secretion than sitosterol. Since biliary sitosterol secretion is preserved, although not elevated in the sitosterolemic mice, this observation suggests that mechanisms other than by Abcg8/sterolin-2 may be responsible for its secretion into bile. [Abstract/Link to Full Text]

Yoon SS, Dambrosia J, Chalela J, Ezzeddine M, Warach S, Haymore J, Davis L, Baird AE
Rising statin use and effect on ischemic stroke outcome.
BMC Med. 2004 Mar 23;24.
BACKGROUND: Statins (3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors) have neuroprotective effects in experimental stroke models and are commonly prescribed in clinical practice. The aim of this study was to determine if patients taking statins before hospital admission for stroke had an improved clinical outcome. METHODS: This was an observational study of 436 patients admitted to the National Institutes of Health Suburban Hospital Stroke Program between July 2000 and December 2002. Self-reported risk factors for stroke were obtained on admission. Stroke severity was determined by the admission National Institutes of Health Stroke Scale score. Good outcome was defined as a Rankin score < 2 at discharge. Statistical analyses used univariate and multivariate logistic regression models. RESULTS: There were 436 patients with a final diagnosis of ischemic stroke; statin data were available for 433 of them. A total of 95/433 (22%) of patients were taking a statin when they were admitted, rising from 16% in 2000 to 26% in 2002. Fifty-one percent of patients taking statins had a good outcome compared to 38% of patients not taking statins (p = 0.03). After adjustment for confounding factors, statin pretreatment was associated with a 2.9 odds (95% CI: 1.2-6.7) of a good outcome at the time of hospital discharge. CONCLUSIONS: The proportion of patients taking statins when they are admitted with stroke is rising rapidly. Statin pretreatment was significantly associated with an improved functional outcome at discharge. This finding could support the early initiation of statin therapy after stroke. [Abstract/Link to Full Text]

MacMaster FP, Kusumakar V
Hippocampal volume in early onset depression.
BMC Med. 2004 Jan 29;22.
BACKGROUND: Abnormalities in limbic structures have been implicated in major depressive disorder (MDD). Although MDD is as common in adolescence as in adulthood, few studies have examined youth near illness onset in order to determine the possible influence of atypical development on the pathophysiology of this disorder. METHODS: Hippocampal volumes were measured in 17 MDD subjects (age = 16.67 +/- 1.83 years [mean +/- SD]; range = 13 - 18 years) and 17 age- and sex-matched healthy controls (16.23 +/- 1.61 years [mean +/- SD]; 13 - 18 years) using magnetic resonance imaging (MRI). RESULTS: An analysis of covariance revealed a significant difference between MDD and control subjects (F = 8.66, df = 1, 29, P = 0.006). This was more strongly localized to the left hippocampus (P = 0.001) than the right hippocampus (P = 0.047). CONCLUSIONS: Our findings provide new evidence of abnormalities in the hippocampus in early onset depression. However, our results should be considered preliminary given the small sample size studied. [Abstract/Link to Full Text]

Papaioannou A, Parkinson W, Cook R, Ferko N, Coker E, Adachi JD
Prediction of falls using a risk assessment tool in the acute care setting.
BMC Med. 2004 Jan 21;21.
BACKGROUND: The British STRATIFY tool was previously developed to predict falls in hospital. Although the tool has several strengths, certain limitations exist which may not allow generalizability to a Canadian setting. Thus, we tested the STRATIFY tool with some modification and re-weighting of items in Canadian hospitals. METHODS: This was a prospective validation cohort study in four acute care medical units of two teaching hospitals in Hamilton, Ontario. In total, 620 patients over the age of 65 years admitted during a 6-month period. Five patient characteristics found to be risk factors for falls in the British STRATIFY study were tested for predictive validity. The characteristics included history of falls, mental impairment, visual impairment, toileting, and dependency in transfers and mobility. Multivariate logistic regression was used to obtain optimal weights for the construction of a risk score. A receiver-operating characteristic curve was generated to show sensitivities and specificities for predicting falls based on different threshold scores for considering patients at high risk. RESULTS: Inter-rater reliability for the weighted risk score indicated very good agreement (inter-class correlation coefficient = 0.78). History of falls, mental impairment, toileting difficulties, and dependency in transfer / mobility significantly predicted fallers. In the multivariate model, mental status was a significant predictor (P < 0.001) while history of falls and transfer / mobility difficulties approached significance (P = 0.089 and P = 0.077 respectively). The logistic regression model led to weights for a risk score on a 30-point scale. A risk score of 9 or more gave a sensitivity of 91% and specificity of 60% for predicting who would fall. CONCLUSION: Good predictive validity for identifying fallers was achieved in a Canadian setting using a simple-to-obtain risk score that can easily be incorporated into practice. [Abstract/Link to Full Text]

Feudtner C, DiGiuseppe DL, Neff JM
Hospital care for children and young adults in the last year of life: a population-based study.
BMC Med. 2003 Dec 23;13.
BACKGROUND: To help design population-based pediatric palliative care services, we sought to describe the hospital care received in the last year of life by children and young adults who died. We also determined the proportion with complex chronic conditions (CCCs) and tested whether the use of hospital services increased as the date of death drew nearer. METHODS: For all deaths occurring under 25 years of age from 1990 to 1996 in Washington State, USA, we linked death certificate information to hospital utilization records and analyzed the timing and duration of hospitalizations and the nature of hospital procedures during the year prior to death. RESULTS: Of the 8 893 deaths, 25 % had CCCs. Among infants with CCCs, 84 % were hospitalized at the time of death and 50 % had been mechanically ventilated during their terminal admission. Among the 458 CCC neonates dying under a week of age, 92% of all days of life were spent in the hospital; among the 172 CCC neonates dying during the second to fourth weeks of life, 85 % of all days of life were spent hospitalized; among the 286 CCC infants dying during the second to twelfth month of life, 41 % of all days of life were spent hospitalized. Among children and young adults with CCCs, 55 % were hospitalized at the time of death, and 19 % had been mechanically ventilated during their terminal admission. For these older patients, the median number of days spent in the hospital during the year preceding death was 18, yet less than a third of this group was hospitalized at any point in time until the last week of their lives. The rate of hospital use increased as death drew near. For subjects who had received hospital care, 44 % had governmental insurance as the source of primary payment. CONCLUSIONS: Infants who died spent a substantial proportion of their lives in hospitals, whereas children and adolescents who died from CCCs predominantly lived outside of the hospital during the last year of life. To serve these patients, pediatric palliative and end-of-life care will have to be provided in an integrated, coordinated manner both in hospitals and home communities. [Abstract/Link to Full Text]

Figueredo A, Zuraw L, Wong RK, Agboola O, Rumble RB, Tandan V
The use of preoperative radiotherapy in the management of patients with clinically resectable rectal cancer: a practice guideline.
BMC Med. 2003 Nov 24;11.
BACKGROUND: This systematic review with meta-analysis was designed to evaluate the literature and to develop recommendations regarding the use of preoperative radiotherapy in the management of patients with resectable rectal cancer. METHODS: The MEDLINE, CANCERLIT and Cochrane Library databases, and abstracts published in the annual proceedings of the American Society of Clinical Oncology and the American Society for Therapeutic Radiology and Oncology were systematically searched for evidence. Relevant reports were reviewed by four members of the Gastrointestinal Cancer Disease Site Group and the references from these reports were searched for additional trials. External review by Ontario practitioners was obtained through a mailed survey. Final approval of the practice guideline report was obtained from the Practice Guidelines Coordinating Committee. RESULTS: Two meta-analyses of preoperative radiotherapy versus surgery alone, nineteen trials that compared preoperative radiotherapy plus surgery to surgery alone, and five trials that compared preoperative radiotherapy to alternative treatments were obtained. Randomized trials demonstrate that preoperative radiotherapy followed by surgery is significantly more effective than surgery alone in preventing local recurrence in patients with resectable rectal cancer and it may also improve survival. A single trial, using surgery with total mesorectal excision, has shown similar benefits in local recurrence. CONCLUSION: For adult patients with clinically resectable rectal cancer we conclude that: Preoperative radiotherapy is an acceptable alternative to the previous practice of postoperative radiotherapy for patients with stage II and III resectable rectal cancer. Both preoperative and postoperative radiotherapy decrease local recurrence but neither improves survival as much as postoperative radiotherapy combined with chemotherapy. Therefore, if preoperative radiotherapy is used, chemotherapy should be added postoperatively to at least patients with stage III disease. [Abstract/Link to Full Text]

Montori VM, Wilczynski NL, Morgan D, Haynes RB
Systematic reviews: a cross-sectional study of location and citation counts.
BMC Med. 2003 Nov 24;12.
BACKGROUND: Systematic reviews summarize all pertinent evidence on a defined health question. They help clinical scientists to direct their research and clinicians to keep updated. Our objective was to determine the extent to which systematic reviews are clustered in a large collection of clinical journals and whether review type (narrative or systematic) affects citation counts. METHODS: We used hand searches of 170 clinical journals in the fields of general internal medicine, primary medical care, nursing, and mental health to identify review articles (year 2000). We defined 'review' as any full text article that was bannered as a review, overview, or meta-analysis in the title or in a section heading, or that indicated in the text that the intention of the authors was to review or summarize the literature on a particular topic. We obtained citation counts for review articles in the five journals that published the most systematic reviews. RESULTS: 11% of the journals concentrated 80% of all systematic reviews. Impact factors were weakly correlated with the publication of systematic reviews (R2 = 0.075, P = 0.0035). There were more citations for systematic reviews (median 26.5, IQR 12 - 56.5) than for narrative reviews (8, 20, P <.0001 for the difference). Systematic reviews had twice as many citations as narrative reviews published in the same journal (95% confidence interval 1.5 - 2.7). CONCLUSIONS: A few clinical journals published most systematic reviews. Authors cited systematic reviews more often than narrative reviews, an indirect endorsement of the 'hierarchy of evidence'. [Abstract/Link to Full Text]

Recent Articles in BMC Complementary and Alternative Medicine

Bell IR, Cunningham V, Caspi O, Meek P, Ferro L
Development and validation of a new global well-being outcomes rating scale for integrative medicine research.
BMC Complement Altern Med. 2004 Jan 15;41.
BACKGROUND: Researchers are finding limitations of currently available disease-focused questionnaire tools for outcome studies in complementary and alternative medicine/integrative medicine (CAM/IM). METHODS: Three substudies investigated the new one-item visual analogue Arizona Integrative Outcomes Scale (AIOS), which assesses self-rated global sense of spiritual, social, mental, emotional, and physical well-being over the past 24 hours and the past month. The first study tested the scale's ability to discriminate unhealthy individuals (n = 50) from healthy individuals (n = 50) in a rehabilitation outpatient clinic sample. The second study examined the concurrent validity of the AIOS by comparing ratings of global well-being to degree of psychological distress as measured by the Brief Symptom Inventory (BSI) in undergraduate college students (N = 458). The third study evaluated the relationships between the AIOS and positively- and negatively-valenced tools (Positive and Negative Affect Scale and the Positive States of Mind Scale) in a different sample of undergraduate students (N = 62). RESULTS: Substudy (i) Rehabilitation patients scored significantly lower than the healthy controls on both forms of the AIOS and a current global health rating. The AIOS 24-hours correlated moderately and significantly with global health (patients r = 0.50; controls r = 0.45). AIOS 1-month correlations with global health were stronger within the controls (patients r = 0.36; controls r = 0.50). Controls (r = 0.64) had a higher correlation between the AIOS 24-hour and 1-month forms than did the patients (r = 0.33), which is consistent with the presumptive improvement in the patients' condition over the previous 30 days in rehabilitation. Substudy (ii) In undergraduate students, AIOS scores were inversely related to distress ratings, as measured by the global severity index on the BSI (rAIOS24h = -0.42, rAIOS1month = -0.40). Substudy (iii) AIOS scores were significantly correlated with positive affect (rAIOS24h = 0.56, rAIOS1month = 0.57) and positive states of mind (rAIOS24h = 0.42, rAIOS1month = 0.45), and inversely correlated with negative affect (rAIOS24h = -0.41, rAIOS1month = -0.59). CONCLUSIONS: The AIOS is able to distinguish relatively sicker from relatively healthier individuals; and correlates in expected directions with a measure of distress and indicators of positive and negative affect and positive states of mind. The AIOS offers a tool for CAM/IM research that extends beyond a disease emphasis. [Abstract/Link to Full Text]

Cuellar N, Aycock T, Cahill B, Ford J
Complementary and alternative medicine (CAM) use by African American (AA) and Caucasian American (CA) older adults in a rural setting: a descriptive, comparative study.
BMC Complement Altern Med. 2003 Nov 18;38.
BACKGROUND: The use of CAM is at an all time high. There is very little research that compares the use of CAM in elders by ethnicity in rural settings. The purpose of the study was to determine if there was a difference between African American and Caucasian American rural elders on use of CAM and self-reported satisfaction with CAM. METHODS: The design was a descriptive, comparative study of 183 elders who reported the number of CAM used and satisfaction with CAM. A convenience sample was recruited through community service organizations in the state of Mississippi. The availability of elders through the support groups, sampling bias, subject effect, and self-report were limitations of the study. RESULTS: The commonest examples of CAM used by rural elders were prayer, vitamins, exercise, meditation, herbs, chiropractic medicine, glucosamine, and music therapy. Significant findings on SES and marital status were calculated. Differences on ethnicity and demographic variables were significant for age, education, and the use of glucosamine. CONCLUSIONS: Health care providers must be aware that elders are using CAM and are satisfied with their use. Identifying different uses of CAM by ethnicity is important for health care practitioners, impacting how health care is provided. [Abstract/Link to Full Text]

Mallick P, Mallick JC, Guha B, Khuda-Bukhsh AR
Ameliorating effect of microdoses of a potentized homeopathic drug, Arsenicum Album, on arsenic-induced toxicity in mice.
BMC Complement Altern Med. 2003 Oct 22;37.
BACKGROUND: Arsenic in groundwater and its accumulation in plants and animals have assumed a menacing proportion in a large part of West Bengal, India and adjoining areas of Bangladesh. Because of the tremendous magnitude of the problem, there seems to be no way to tackle the problem overnight. Efforts to provide arsenic free water to the millions of people living in these dreaded zones are being made, but are awfully inadequate. In our quest for finding out an easy, safe and affordable means to combat this problem, a homeopathic drug, Arsenicum Album-30, appears to yield promising results in mice. The relative efficacies of two micro doses of this drug, namely, Arsenicum Album-30 and Arsenicum Album-200, in combating arsenic toxicity have been determined in the present study on the basis of some accepted biochemical protocols. METHODS: Mice were divided into different sets of control (both positive and negative) and treated series (As-intoxicated, As-intoxicated plus drug-fed). Alanine amino transferase (ALT) and aspartate amino transferase (AST) activities and reduced glutathione (GSH) level in liver and blood were analyzed in the different series of mice at six different fixation intervals. RESULTS: Both Arsenicum Album-30 and Arsenicum Album-200 ameliorated arsenic-induced toxicity to a considerable extent as compared to various controls. CONCLUSIONS: The results lend further support to our earlier views that microdoses of potentized Arsenicum Album are capable of combating arsenic intoxication in mice, and thus are strong candidates for possible use in human subjects in arsenic contaminated areas under medical supervision. [Abstract/Link to Full Text]

Tajuddin S, Latif A, Qasmi IA
Aphrodisiac activity of 50% ethanolic extracts of Myristica fragrans Houtt. (nutmeg) and Syzygium aromaticum (L) Merr. & Perry. (clove) in male mice: a comparative study.
BMC Complement Altern Med. 2003 Oct 20;36.
BACKGROUND: Spices are considered as sexual invigorators in the Unani System of Medicine. In order to explore the sexual function improving effect of Myristica fragrans Houtt. (nutmeg) and Syzygium aromaticum (L) Merr. & Perry. (clove) an experimental study was conducted in normal male mice. METHODS: The extracts (50% ethanolic) of nutmeg and clove were administered (500 mg/kg; p.o.) to different groups of male Swiss mice. Mounting behaviour, mating performance, and general short term toxicity of the test drugs were determined and compared with the standard drug Penegra (Sildenafil citrate). RESULTS: The extracts of the nutmeg and clove were found to stimulate the mounting behaviour of male mice, and also to significantly increase their mating performance. The drugs were devoid of any conspicuous general short term toxicity. CONCLUSION: The extracts (50% ethanolic) of nutmeg and clove enhanced the sexual behaviour of male mice. [Abstract/Link to Full Text]

Nammi S, Gudavalli R, Babu BS, Lodagala DS, Boini KM
Possible mechanisms of hypotension produced 70% alcoholic extract of Terminalia arjuna (L.) in anaesthetized dogs.
BMC Complement Altern Med. 2003 Oct 16;35.
BACKGROUND: The bark of Terminalia arjuna L. (Combretaceae) is used in Ayurveda since ancient times for the treatment of cardiac disorders. Previous laboratory investigations have demonstrated the use of the bark in cardiovascular complications. The present study was aimed to find the effect of 70% alcoholic extract of Terminalia arjuna on anaesthetized dog blood pressure and probable site of action. METHODS: Six dogs were anaesthetized with intraperitoneal injection of thiopental sodium and the blood pressure of each dog (n = 6) was measured from the left common carotid artery connected to a mercury manometer on kymograph. The femoral vein was cannulated for administration of drug solutions. The extract of T. arjuna (dissolved in propylene glycol) in the dose range of 5 to 15 mg/kg were administered intravenously in a pilot study and the dose (6 mg/kg) which produced appreciable hypotension was selected for further studies. RESULTS: Intravenous administration of T. arjuna produced dose-dependent hypotension in anaesthetized dogs. The hypotension produced by 6 mg/kg dose of the extract was blocked by propranolol but not by atropine or mepyramine maleate. This indicates that muscarinic or histaminergic mechanisms are not likely to be involved in the hypotension produced by the extract. The blockade by propranolol of the hypotension produced by T. arjuna indicates that the extract might contain active compound(s) possessing adrenergic beta2-receptor agonist action and/or that act directly on the heart muscle. CONCLUSION: The results indicated the likely involvement of peripheral mechanism for hypotension produced by the 70% alcoholic extract of Terminalia arjuna and lends support for the claims of its traditional usage in cardiovascular disorders. [Abstract/Link to Full Text]

Nammi S, Boini MK, Lodagala SD, Behara RB
The juice of fresh leaves of Catharanthus roseus Linn. reduces blood glucose in normal and alloxan diabetic rabbits.
BMC Complement Altern Med. 2003 Sep 2;34.
BACKGROUND: The leaf juice or water decoction of Catharanthus roseus L. (Apocyanaceae) is used as a folk medicine for the treatment of diabetes all over the world. In the present investigation, the leaf juice of C. roseus has been evaluated for its hypoglycemic activity in normal and alloxan-induced diabetic rabbits. METHODS: The blood glucose lowering activity of the leaf juice was studied in normal and alloxan-induced (100 mg/kg, i.v.) diabetic rabbits, after oral administration at doses of 0.5, 0.75 and 1.0 ml/kg body weight. Blood samples were collected from the marginal ear vein before and also at 4, 6, 8, 10, 12, 16, 18, 20 & 24 h after drug administration and blood glucose was analyzed by Nelson-Somogyi's method using a visible spectrophotometer. The data was compared statistically by using Student's t-test. RESULTS: The leaf juice of C. roseus produced dose-dependent reduction in blood glucose of both normal and diabetic rabbits and comparable with that of the standard drug, glibenclamide. The results indicate a prolonged action in reduction of blood glucose by C. roseus and the mode of action of the active compound(s) of C. roseus is probably mediated through enhance secretion of insulin from the beta-cells of Langerhans or through extrapancreatic mechanism. CONCLUSIONS: The present study clearly indicated a significant antidiabetic activity with the leaf juice of Catharanthus roseus and supports the traditional usage of the fresh leaves by Ayurvedic physicians for the control of diabetes. [Abstract/Link to Full Text]

Murphy LS, Reinsch S, Najm WI, Dickerson VM, Seffinger MA, Adams A, Mishra SI
Searching biomedical databases on complementary medicine: the use of controlled vocabulary among authors, indexers and investigators.
BMC Complement Altern Med. 2003 Jul 7;33.
BACKGROUND: The optimal retrieval of a literature search in biomedicine depends on the appropriate use of Medical Subject Headings (MeSH), descriptors and keywords among authors and indexers. We hypothesized that authors, investigators and indexers in four biomedical databases are not consistent in their use of terminology in Complementary and Alternative Medicine (CAM). METHODS: Based on a research question addressing the validity of spinal palpation for the diagnosis of neuromuscular dysfunction, we developed four search concepts with their respective controlled vocabulary and key terms. We calculated the frequency of MeSH, descriptors, and keywords used by authors in titles and abstracts in comparison to standard practices in semantic and analytic indexing in MEDLINE, MANTIS, CINAHL, and Web of Science. RESULTS: Multiple searches resulted in the final selection of 38 relevant studies that were indexed at least in one of the four selected databases. Of the four search concepts, validity showed the greatest inconsistency in terminology among authors, indexers and investigators. The use of spinal terms showed the greatest consistency. Of the 22 neuromuscular dysfunction terms provided by the investigators, 11 were not contained in the controlled vocabulary and six were never used by authors or indexers. Most authors did not seem familiar with the controlled vocabulary for validity in the area of neuromuscular dysfunction. Recently, standard glossaries have been developed to assist in the research development of manual medicine. CONCLUSIONS: Searching biomedical databases for CAM is challenging due to inconsistent use of controlled vocabulary and indexing procedures in different databases. A standard terminology should be used by investigators in conducting their search strategies and authors when writing titles, abstracts and submitting keywords for publications. [Abstract/Link to Full Text]

van Blitterswijk WJ, van de Nes JC, Wuisman PI
Glucosamine and chondroitin sulfate supplementation to treat symptomatic disc degeneration: biochemical rationale and case report.
BMC Complement Altern Med. 2003 Jun 10;32.
BACKGROUND: Glucosamine and chondroitin sulfate preparations are widely used as food supplements against osteoarthritis, but critics are skeptical about their efficacy, because of the lack of convincing clinical trials and a reasonable scientific rationale for the use of these nutraceuticals. Most trials were on osteoarthritis of the knee, while virtually no documentation exists on spinal disc degeneration. The purpose of this article is to highlight the potential of these food additives against cartilage degeneration in general, and against symptomatic spinal disc degeneration in particular, as is illustrated by a case report. The water content of the intervertebral disc is a reliable measure of its degeneration/ regeneration status, and can be objectively determined by Magnetic Resonance Imaging (MRI) signals. CASE PRESENTATION: Oral intake of glucosamine and chondroitin sulfate for two years associated with disk recovery (brightening of MRI signal) in a case of symptomatic spinal disc degeneration. We provide a biochemical explanation for the possible efficacy of these nutraceuticals. They are bioavailable to cartilage chondrocytes, may stimulate the biosynthesis and inhibit the breakdown of their extracellular matrix proteoglycans. CONCLUSION: The case suggests that long-term glucosamine and chondroitin sulfate intake may counteract symptomatic spinal disc degeneration, particularly at an early stage. However, definite proof requires well-conducted clinical trials with these food supplements, in which disc de-/regeneration can be objectively determined by MRI. A number of biochemical reasons (that mechanistically need to be further resolved) explain why these agents may have cartilage structure- and symptom-modifying effects, suggesting their therapeutic efficacy against osteoarthritis in general. [Abstract/Link to Full Text]

Najm WI, Seffinger MA, Mishra SI, Dickerson VM, Adams A, Reinsch S, Murphy LS, Goodman AF
Content validity of manual spinal palpatory exams - A systematic review.
BMC Complement Altern Med. 2003 May 7;31.
BACKGROUND: Many health care professionals use spinal palpatory exams as a primary and well-accepted part of the evaluation of spinal pathology. However, few studies have explored the validity of spinal palpatory exams. To evaluate the status of the current scientific evidence, we conducted a systematic review to assess the content validity of spinal palpatory tests used to identify spinal neuro-musculoskeletal dysfunction. METHODS: Review of eleven databases and a hand search of peer-reviewed literature, published between 1965-2002, was undertaken. Two blinded reviewers abstracted pertinent data from the retrieved papers, using a specially developed quality-scoring instrument. Five papers met the inclusion/exclusion criteria. RESULTS: Three of the five papers included in the review explored the content validity of motion tests. Two of these papers focused on identifying the level of fixation (decreased mobility) and one focused on range of motion. All three studies used a mechanical model as a reference standard. Two of the five papers included in the review explored the validity of pain assessment using the visual analogue scale or the subjects' own report as reference standards. Overall the sensitivity of studies looking at range of motion tests and pain varied greatly. Poor sensitivity was reported for range of motion studies regardless of the examiner's experience. A slightly better sensitivity (82%) was reported in one study that examined cervical pain. CONCLUSIONS: The lack of acceptable reference standards may have contributed to the weak sensitivity findings. Given the importance of spinal palpatory tests as part of the spinal evaluation and treatment plan, effort is required by all involved disciplines to create well-designed and implemented studies in this area. [Abstract/Link to Full Text]

Siddiqui TA, Zafar S, Iqbal N, Nadeem A, Zaidi Z, Alavi SH
Effect of Kohl-Chikni Dawa - a compound ophthalmic formulation of Unani medicine on naphthalene-induced cataracts in rats.
BMC Complement Altern Med. 2002 Dec 29;213.
BACKGROUND: Cataracts are the leading cause of blindness worldwide, accounting for 13-27% of cases. Kohl-Chikni Dawa (KCD) is reputed for its beneficial effects in the treatment of premature cataracts. However, its efficacy is yet to be tested. To investigate the rationality of the therapeutic use of Kohl-Chikni Dawa (KCD) in Unani medicine. METHODS: The effect of Kohl-Chikni Dawa eye drops on naphthalene-induced cataracts in rats was investigated by slit-lamp biomicroscopic analysis. The normal group of experimental animals was administered with mineral oil (orally), while other groups were given naphthalene (orally) along with local application of KCD eye drops (once and twice daily), placebo and distilled water (twice daily). Initial morphological changes of the lenses were observed twice a week for two weeks, and thereafter once a week for four weeks. RESULTS: Local application of KCD (twice daily) caused significant reduction in the lens opacification after 2 to 4 weeks of naphthalene administration. CONCLUSION: KCD eye drops may have the potential to delay progression of naphthalene-induced cataracts in rats. [Abstract/Link to Full Text]

Lee JS, Mamo J, Ho N, Pal S
The effect of Puerariae radix on lipoprotein metabolism in liver and intestinal cells.
BMC Complement Altern Med. 2002 Dec 16;212.
BACKGROUND: Animal studies investigating the beneficial effects of Puerariae radix on cardiovascular disease have suggested this plant possesses anti-diabetic and lipid lowering properties. However, the exact mechanism by which Puerariae radix affects lipid metabolism is currently unknown. The aim of this study was to investigate the effect of the water extract of Puerariae radix on the secretion of VLDL and chylomicrons from HepG2 liver cells and CaCo2 cells, respectively, in humans. METHODS: The amount of apoB100 (a protein marker for VLDL) and apoB48 (a protein marker for chylomicrons) in cells and media were quantified by Western Blotting and enhanced chemiluminescence (ECL). Total, free and esterified cholesterol concentrations were measured by gas liquid chromatography. RESULTS: Treatment of cells with water extract of Puerariae radix significantly decreased apoB100 production and secretion from HepG2 cells up to 66% in a dose dependent manner. The intracellular total cholesterol and free cholesterol concentration in HepG2 cells also decreased with increasing concentration of the Puerariae radix. In contrast, water extract of Puerariae radix attenuated apoB48 concentrations in cells, but not apoB48 secretion from CaCo2 enterocytes. CONCLUSIONS: Collectively, our findings suggest that the water extract of Puerariae radix attenuates the hepatic lipoprotein production and secretion. Our present cell culture findings may explain why circulating VLDL and LDL levels were attenuated in animals supplemented with Puerariae radix. Since decreasing the production and secretion of atherogenic lipoproteins decreases the risk of development of cardiovascular disease, diets supplemented with radix may provide a safe and effective beneficial cardioprotective effects in humans. [Abstract/Link to Full Text]

Cooper R, Dragar C, Elliot K, Fitton JH, Godwin J, Thompson K
GFS, a preparation of Tasmanian Undaria pinnatifida is associated with healing and inhibition of reactivation of Herpes.
BMC Complement Altern Med. 2002 Nov 20;211.
BACKGROUND: We sought to assess whether GFS, a proprietary preparation of Tasmanian Undaria pinnatifida, has effects on healing or re-emergence of Herpetic infections, and additionally, to assess effects of GFS in vitro. Undaria is the most commonly eaten seaweed in Japan, and contains sulphated polyanions and other components with potential anti-viral activity. Herpes simplex virus type 1 (HSV-1) infections have lower reactivation rates and Herpes type 2 (HSV-2) infections have lower incidence in Japan than in the west. METHODS: Patients with active (15 subjects) or latent (6 subjects) Herpetic infections (HSV-1, 2, EBV, Zoster) were monitored for response to ingestion of GFS. GFS extract was tested in vitro for human T cell mitogenicity and anti-Herpes activity. RESULTS: Ingestion of GFS was associated with increased healing rates in patients with active infections. In addition, patients with latent infection remained asymptomatic whilst ingesting GFS. GFS extract inhibited Herpes viruses in vitro and was mitogenic to human T cells in vitro. CONCLUSIONS: Ingestion of GFS has inhibitory effects on reactivation and is associated with increased rate of healing after Herpetic outbreaks. GFS extract potently inhibited Herpes virus in vitro, and had mitogenic effects on human T cells. [Abstract/Link to Full Text]

Moreland N, La Grange L, Montoya R
Impact of in utero exposure to EtOH on corpus callosum development and paw preference in rats: protective effects of silymarin.
BMC Complement Altern Med. 2002 Nov 11;210.
BACKGROUND: Using a rat model we have found that the bioflavonoid silymarin (SY) ameliorates some of the negative consequences of in utero exposure to ethanol (EtOH). In the current study our aim was to determine if laterality preference and corpus callosum development were altered in rat offspring whose mothers were provided with a concomitant administration of SY with EtOH throughout gestation. METHODS: We provided pregnant Fisher/344 rats with liquid diets containing 35% ethanol derived calories (EDC) throughout the gestational period. A silymarin/phospholipid compound containing 29.8% silybin was co administered with EtOH to a separate experimental group. We tested the offspring for laterality preference at age 12 weeks. After testing the rats were sacrificed and their brains perfused for later corpus callosum extraction. RESULTS: We observed incomplete development of the splenium in the EtOH-only offspring. Callosal development was complete in all other treatment groups. Rats from the EtOH-only group displayed a left paw preference; whereas control rats were evenly divided between right and left paw preference. Inexplicably both SY groups were largely right paw preferring. CONCLUSIONS: The addition of SY to the EtOH liquid diet did confer some ameliorative effects upon the developing fetal rat brain. [Abstract/Link to Full Text]

Tanaka TH, Leisman G, Mori H, Nishijo K
The effect of massage on localized lumbar muscle fatigue.
BMC Complement Altern Med. 2002 Oct 14;29.
BACKGROUND: There is not enough evidence to support the efficacy of massage for muscle fatigue despite wide utilization of the modality in various clinical settings. This study investigated the influence of massage application on localized back muscle fatigue. METHODS: Twenty-nine healthy subjects participated in two experimental sessions (massage and rest conditions). On each test day, subjects were asked to lie in the prone position on a treatment table and perform sustained back extension for 90 seconds. Subjects then either received massage on the lumbar region or rested for a 5 minute duration, then repeated the back extension movement. The median frequency (MDF), mean power frequency (MNF), and root mean square (RMS) amplitude of electromyographic signals during the 90 second sustained lumbar muscle contraction were analyzed. The subjective feeling of fatigue was then evaluated using the Visual Analogue Scale (VAS). RESULTS: MDF and MNF significantly declined with time under all conditions. There was no significant difference in MDF, MNF or RMS value change between before and after massage, or between rest and massage conditions. There was a significant increase in fatigue VAS at the end of the 2nd back extension with rest condition. There was a significant difference in fatigue VAS change between massage and rest condition. CONCLUSIONS: A significant difference was observed between massage and rest condition on VAS for muscle fatigue. On EMG analysis, there were no significant differences to conclude that massage stimulation influenced the myoelectrical muscle fatigue, which is associated with metabolic and electrical changes. [Abstract/Link to Full Text]

Shen J, Andersen R, Albert PS, Wenger N, Glaspy J, Cole M, Shekelle P
Use of complementary/alternative therapies by women with advanced-stage breast cancer.
BMC Complement Altern Med. 2002 Aug 13;28.
BACKGROUND: This study sought to describe the pattern of complementary/alternative medicine (CAM) use among a group of patients with advanced breast cancer, to examine the main reasons for their CAM use, to identify patient's information sources and their communication pattern with their physicians. METHODS: Face-to-face structured interviews of patients with advanced-stage breast cancer at a comprehensive oncology center. RESULTS: Seventy three percent of patients used CAM; relaxation/meditative techniques and herbal medicine were the most common. The most commonly cited primary reason for CAM use was to boost the immune system, the second, to treat cancer; however these reasons varied depending on specific CAM therapy. Friends or family members and mass media were common primary information source's about CAM. CONCLUSIONS: A high proportion of advanced-stage breast cancer patients used CAM. Discussion with doctors was high for ingested products. Mass media was a prominent source of patient information. Credible sources of CAM information for patients and physicians are needed. [Abstract/Link to Full Text]

Blackmer J, Jefromova L
The use of alternative therapies in the Saskatchewan stroke rehabilitation population.
BMC Complement Altern Med. 2002 Jul 2;27.
BACKGROUND: Many patients use alternative therapies. The purpose of this study was to determine the percentage of stroke rehabilitation patients in Saskatchewan using alternative therapies, whether patients found these therapies effective in alleviating stroke-related symptoms, how often those patients who used alternative therapies discuss this fact with their primary care doctor and the main reason why patients might not do so. METHODS: Telephone questionnaire surveys were conducted with 117 patients who had suffered a stroke and undergone inpatient or outpatient rehabilitation at Saskatoon City Hospital. RESULTS: The study revealed that 26.5% of 117 stroke rehabilitation patients visited alternative practitioners at least once or used some form of unconventional therapy. Only 16.1% of patients found that alternative therapy made them feel much better. Of those who used alternative therapy, 61.3% did not discuss this fact with their primary physician. Many of the respondents (47.3%) who did not inform their physician stated that they did not see the necessity of talking about these treatments and 21.1% did not discuss the issue with their physician because they felt that he or she might disapprove of alternative therapies. CONCLUSION: A relatively small percentage of stroke patients found alternative therapies beneficial. Doctors should be aware that a significant number of patients will try alternative treatment without discussion with their primary care physician or specialist. The current study suggests that after completing routine questioning, doctors should also ask their patients about their use of alternative therapies and, when appropriate, review issues of safety and efficacy. [Abstract/Link to Full Text]

Rahman S, Ali Khan R, Kumar A
Experimental study of the morphine de-addiction properties of Delphinium denudatum Wall.
BMC Complement Altern Med. 2002 May 29;26.
BACKGROUND: Our aim was to explore the de-addiction properties of Delphinium denudatum Wall. in morphine dependent rats. METHODS: Charles Foster male albino rats were made morphine dependent by injecting morphine sulphate in increasing doses twice a day for 7 days. The spontaneous withdrawal signs observed 12 h after the last dose were quantified by the 'counted' and 'checked' signs. The drug (alcoholic extract of Delphinium denudatum) was administered p.o. in different regimen: a) single dose (700 mg/kg) 10 h before the first dose of morphine, b) single dose (700 mg/kg) 10 h after the last dose of morphine, c) multiple doses (350 mg/kg) along with morphine twice a day for 7 days. RESULT: Administration of Delphinium denudatum extract caused significant reduction in the frequency of counted signs as well as the presence of checked signs of morphine withdrawal. The maximum reduction was observed in regimen 'b' followed by regimen 'c' and 'a'. CONCLUSION: Delphinium denudatum Wall. significantly reduces the aggregate scores for all parameters in morphine withdrawal syndrome by central action and thus may prove to be an alternative remedy in morphine de-addiction. [Abstract/Link to Full Text]

Hollyer T, Boon H, Georgousis A, Smith M, Einarson A
The use of CAM by women suffering from nausea and vomiting during pregnancy.
BMC Complement Altern Med. 2002 May 17;25.
BACKGROUND: Nausea and vomiting during pregnancy (NVP) affects two-thirds of pregnant women to varying degrees and over the years many modalities have been used to try to alleviate this often debilitating condition. There is a paucity of information in the literature about the use or efficacy of complementary and alternative medicine (CAM) for the treatment of this condition that affects so many women. Our primary objective was to examine the prevalence of CAM usage by women suffering from NVP. Our secondary objective was to ascertain if women had any supervision in the use of these treatments. METHODS: Women who called The Motherisk NVP helpline, were asked after the counseling session to complete a questionnaire, which included demographic data as well as information about their CAM use. RESULTS: Seventy women completed the questionnaire. 61% reported using CAM therapies, of which the three most popular were: ginger, vitamin B6 and acupressure. 21% of those who reported using CAM, had consulted CAM practitioners, 8% their physicians or pharmacists and 71% discussed the usage with family, friends and other allied health professionals. Women who did not use CAM stated they would probably use these modalities if there was more information about the safety in pregnancy. CONCLUSION: Pregnant women with NVP are mirroring the trend in the general population of the use of CAM. They are also using CAM therapies with little supervision from practitioners experienced in the use of these modalities. [Abstract/Link to Full Text]

Biswas SJ, Khuda-Bukhsh AR
Effect of a homeopathic drug, Chelidonium, in amelioration of p-DAB induced hepatocarcinogenesis in mice.
BMC Complement Altern Med. 2002 Apr 10;24.
BACKGROUND: Crude extracts of Chelidonium majus, and also purified compounds derived from crude extracts of this plant, have been reported to exhibit anti-viral, anti-inflammatory, anti-tumor and anti-microbial properties both in vitro and in vivo. Chelidonium is a homeopathic drug routinely used against various liver disorders including cancer in humans. Two potencies of Chelidonium (Ch-30, Ch-200) have been tested for their possible anti-tumor and enzyme modulating activities in liver and anti-clastogenic effects during p-DAB-induced hepatocarcinogenesis in mice compared to suitable controls. METHODS: Several cytogenetic and enzymatic protocols were used at three fixation intervals; at 60 days, 90 days and 120 days of treatment. Different sets of healthy mice were fed: i) hepatocarcinogen, p-DAB plus phenobarbital (PB), ii) only PB, iii) neither p-DAB nor PB (normal control). One set of mice fed with p-DAB plus PB was also fed Ch-30 (iv) and another set Ch-200 (v). All standard currently used methods were adopted for cytogenetical preparations and for the enzyme assays. RESULTS: All group (i) mice developed tumors in liver at all fixation intervals, while none of group (ii) and (iii) mice developed any tumors. About 40% mice in group (iv) and group (v) did not show tumor nodules in their liver. Feeding of Chelidonium to group (iv) and (v) mice reduced genotoxic effects to a significant extent (p < 0.05 to p < 0.001). CONCLUSION: The homeopathic drug Chelidonium exhibited anti-tumor and anti-genotoxic activities and also favorably modulated activities of some marker enzymes. Microdoses of Chelidonium may be effectively used in combating liver cancer. [Abstract/Link to Full Text]

Cernáková M, Kost'álová D, Kettmann V, Plodová M, Tóth J, Drímal J
Potential antimutagenic activity of berberine, a constituent of Mahonia aquifolium.
BMC Complement Altern Med. 2002 Feb 19;22.
BACKGROUND: As part of a study aimed at developing new pharmaceutical products from natural resources, the purpose of this research was twofold: (1) to fractionate crude extracts from the bark of Mahonia aquifolium and (2) to evaluate the strength of the antimutagenic activity of the separate components against one of the common direct-acting chemical mutagens. METHODS: The antimutagenic potency was evaluated against acridine orange (AO) by using Euglena gracilis as an eukaryotic test model, based on the ability of the test compound/fraction to prevent the mutagen-induced damage of chloroplast DNA. RESULTS: It was found that the antimutagenicity of the crude Mahonia extract resides in both bis-benzylisoquinoline (BBI) and protoberberine alkaloid fractions but only the protoberberine derivatives, jatrorrhizine and berberine, showed significant concentration-dependent inhibitory effect against the AO-induced chloroplast mutagenesis of E. gracilis. Especially berberine elicited, at a very low dose, remarkable suppression of the AO-induced mutagenicity, its antimutagenic potency being almost three orders of magnitude higher when compared to its close analogue, jatrorrhizine. Possible mechanisms of the antimutagenic action are discussed in terms of recent literature data. While the potent antimutagenic activity of the protoberberines most likely results from the inhibition of DNA topoisomerase I, the actual mechanism(s) for the BBI alkaloids is hard to be identified. CONCLUSIONS: Taken together, the results indicate that berberine possesses promising antimutagenic/anticarcinogenic potential that is worth to be investigated further. [Abstract/Link to Full Text]

Sangdee C, Teekachunhatean S, Sananpanich K, Sugandhavesa N, Chiewchantanakit S, Pojchamarnwiputh S, Jayasvasti S
Electroacupuncture versus diclofenac in symptomatic treatment of osteoarthritis of the knee: a randomized controlled trial.
BMC Complement Altern Med. 2002 Mar 21;23.
BACKGROUND: The purpose of this study was to compare the efficacy of electroacupuncture (EA), diclofenac and their combination in symptomatic treatment of osteoarthritis (OA) of the knee. METHODS: This study was a randomized, single-blind, placebo controlled trial. The 193 out-patients with OA of the knee were randomized into four groups: placebo, diclofenac, EA and combined (diclofenac plus EA). Paracetamol tablets were prescribed as a rescue analgesic during the study. The patients were evaluated after a run-in period of one week (week 0) and again at the end of the study (week 4). The clinical assessments included the amount of paracetamol taken/week, visual analog scale (VAS), Western Ontario and McMaster Universities (WOMAC) OA Index, Lequesne's functional index, 50 feet-walk time, and the orthopedist's and patient's opinion of change. RESULTS: One hundred and eighty six patients completed the study. The improvement of symptoms (reduction in mean changes) in most outcome parameters was greatest in the EA group. The proportions of responders and patients with an overall opinion of "much better" were also greatest in the EA group. The improvement in VAS was significantly different between the EA and placebo group as well as the EA and diclofenac group. The improvement in Lequesne's functional index also differed significantly between the EA and placebo group. In addition, there was a significant improvement in WOMAC pain index between the combined and placebo group. CONCLUSION: EA is significantly more effective than placebo and diclofenac in the symptomatic treatment of OA of the knee in some circumstances. However, the combination of EA and diclofenac treatment was no more effective than EA treatment alone. [Abstract/Link to Full Text]

St-Onge MP, Farnworth ER, Savard T, Chabot D, Mafu A, Jones PJ
Kefir consumption does not alter plasma lipid levels or cholesterol fractional synthesis rates relative to milk in hyperlipidemic men: a randomized controlled trial [ISRCTN10820810].
BMC Complement Altern Med. 2002;21.
BACKGROUND: Fermented milk products have been shown to affect serum cholesterol concentrations in humans. Kefir, a fermented milk product, has been traditionally consumed for its potential health benefits but has to date not been studied for its hypocholesterolemic properties. METHODS: Thirteen healthy mildly hypercholesterolemic male subjects consumed a dairy supplement in randomized crossover trial for 2 periods of 4 wk each. Subjects were blinded to the dairy supplement consumed. Blood samples were collected at baseline and after 4 wk of supplementation for measurement of plasma total, low-density lipoprotein, and high-density lipoprotein cholesterol and triglyceride concentrations, as well as fatty acid profile and cholesterol synthesis rate. Fecal samples were collected at baseline and after 2 and 4 wk of supplementation for determination of fecal short chain fatty acid level and bacterial content. RESULTS: Kefir had no effect on total cholesterol, low-density lipoprotein cholesterol, high-density lipoprotein cholesterol or triglyceride concentrations nor on cholesterol fractional synthesis rates after 4 wk of supplementation. No significant change on plasma fatty acid levels was observed with diet. However, both kefir and milk increased (p < 0.05) fecal isobutyric, isovaleric and propionic acids as well as the total amount of fecal short chain fatty acids. Kefir supplementation resulted in increased fecal bacterial content in the majority of the subjects. CONCLUSIONS: Since kefir consumption did not result in lowered plasma lipid concentrations, the results of this study do not support consumption of kefir as a cholesterol-lowering agent. [Abstract/Link to Full Text]

Jonas WB, Anderson RL, Crawford CC, Lyons JS
A systematic review of the quality of homeopathic clinical trials.
BMC Complement Altern Med. 2001;112.
BACKGROUND: While a number of reviews of homeopathic clinical trials have been done, all have used methods dependent on allopathic diagnostic classifications foreign to homeopathic practice. In addition, no review has used established and validated quality criteria allowing direct comparison of the allopathic and homeopathic literature. METHODS: In a systematic review, we compared the quality of clinical-trial research in homeopathy to a sample of research on conventional therapies using a validated and system-neutral approach. All clinical trials on homeopathic treatments with parallel treatment groups published between 1945-1995 in English were selected. All were evaluated with an established set of 33 validity criteria previously validated on a broad range of health interventions across differing medical systems. Criteria covered statistical conclusion, internal, construct and external validity. Reliability of criteria application is greater than 0.95. RESULTS: 59 studies met the inclusion criteria. Of these, 79% were from peer-reviewed journals, 29% used a placebo control, 51% used random assignment, and 86% failed to consider potentially confounding variables. The main validity problems were in measurement where 96% did not report the proportion of subjects screened, and 64% did not report attrition rate. 17% of subjects dropped out in studies where this was reported. There was practically no replication of or overlap in the conditions studied and most studies were relatively small and done at a single-site. Compared to research on conventional therapies the overall quality of studies in homeopathy was worse and only slightly improved in more recent years. CONCLUSIONS: Clinical homeopathic research is clearly in its infancy with most studies using poor sampling and measurement techniques, few subjects, single sites and no replication. Many of these problems are correctable even within a "holistic" paradigm given sufficient research expertise, support and methods. [Abstract/Link to Full Text]

Miller MJ, Angeles FM, Reuter BK, Bobrowski P, Sandoval M
Dietary antioxidants protect gut epithelial cells from oxidant-induced apoptosis.
BMC Complement Altern Med. 2001;111.
BACKGROUND: The potential of ascorbic acid and two botanical decoctions, green tea and cat's claw, to limit cell death in response to oxidants were evaluated in vitro. METHODS: Cultured human gastric epithelial cells (AGS) or murine small intestinal epithelial cells (IEC-18) were exposed to oxidants - DPPH (3 microM), H2O2 (50 microM), peroxynitrite (300 microM) - followed by incubation for 24 hours, with antioxidants (10 microg/ml) administered as a 1 hour pretreatment. Cell number (MTT assay) and death via apoptosis or necrosis (ELISA, LDH release) was determined. The direct interactions between antioxidants and DPPH (100 microM) or H2O2 (50 microM) were evaluated by spectroscopy. RESULTS: The decoctions did not interact with H2O2, but quenched DPPH although less effectively than vitamin C. In contrast, vitamin C was significantly less effective in protecting human gastric epithelial cells (AGS) from apoptosis induced by DPPH, peroxynitrite and H2O2 (P < 0.001). Green tea and cat's claw were equally protective against peroxynitrite and H2O2, but green tea was more effective than cat's claw in reducing DPPH-induced apoptosis (P < 0.01). Necrotic cell death was marginally evident at these low concentrations of peroxynitrite and H2O2, and was attenuated both by cat's claw and green tea (P < 0.01). In IEC-18 cells, all antioxidants were equally effective as anti-apoptotic agents. CONCLUSIONS: These results indicate that dietary antioxidants can limit epithelial cell death in response to oxidant stress. In the case of green tea and cat's claw, the cytoprotective response exceed their inherent ability to interact with the injurious oxidant, suggestive of actions on intracellular pathways regulating cell death. [Abstract/Link to Full Text]

Datta SS, Mallick PP, Rahman Khuda-Bukhsh AA
Comparative efficacy of two microdoses of a potentized homoeopathic drug, Cadmium Sulphoricum, in reducing genotoxic effects produced by cadmium chloride in mice: a time course study.
BMC Complement Altern Med. 2001;19.
BACKGROUND: Cadmium poisoning in the environment has assumed an alarming problem in recent years. Effective antimutagenic agents which can reverse or combat cadmium induced genotoxicity in mice have not yet been reported. Therefore, in the present study, following the homeopathic principle of "like cures like", we tested the efficacy of two potencies of a homeopathic drug, Cadmium Sulphoricum (Cad Sulph), in reducing the genotoxic effects of Cadmium chloride in mice.Another objective was to determine the relative efficacy of three administrative modes, i.e. pre-, post- and combined pre and post-feeding of the homeopathic drugs. For this, healthy mice, Mus musculus, were intraperitoneally injected with 0.008% solution of CdCl2 @ 1 ml/100 gm of body wt (i.e. 0.8 mcg/gm of bw), and assessed for the genotoxic effects through such studies as chromosome aberrations (CA), micronucleated erythrocytes (MNE), mitotic index (MI) and sperm head anomaly (SHA), keeping suitable succussed alcohol fed (positive) and CdCl2 untreated normal (negative) controls. The CdCl2 treated mice were divided into 3 subgroups, which were orally administered with the drug prior to, after and both prior to and after injection of CdCl2 at specific fixation intervals and their genotoxic effects were analyzed. RESULTS: While the CA, MNE and SHA were reduced in the drug fed series as compared to their respective controls, the MI showed an apparent increase. The combined pre- and post-feeding of Cad Sulph showed maximum reduction of the genotoxic effects. CONCLUSIONS: Both Cad Sulph-30 and 200 were able to combat cadmium induced genotoxic effects in mice and that combined pre- and post-feeding mode of administration was found to be most effective in reducing the genotoxic effect of CdCl2 followed by the post-feeding mode. [Abstract/Link to Full Text]

Lans C, Harper T, Georges K, Bridgewater E
Medicinal and ethnoveterinary remedies of hunters in Trinidad.
BMC Complement Altern Med. 2001;110.
BACKGROUND: Ethnomedicines are used by hunters for themselves and their hunting dogs in Trinidad. Plants are used for snakebites, scorpion stings, for injuries and mange of dogs and to facilitate hunting success. RESULTS: Plants used include Piper hispidum, Pithecelobium unguis-cati, Bauhinia excisa, Bauhinia cumanensis, Cecropia peltata, Aframomum melegueta, Aristolochia rugosa, Aristolochia trilobata, Jatropha curcas, Jatropha gossypifolia, Nicotiana tabacum, Vernonia scorpioides, Petiveria alliacea, Renealmia alpinia, Justicia secunda, Phyllanthus urinaria,Phyllanthus niruri,Momordica charantia, Xiphidium caeruleum, Ottonia ovata, Lepianthes peltata, Capsicum frutescens, Costus scaber, Dendropanax arboreus, Siparuma guianensis, Syngonium podophyllum, Monstera dubia, Solanum species, Eclipta prostrata, Spiranthes acaulis, Croton gossypifolius, Barleria lupulina, Cola nitida, Acrocomia ierensis (tentative ID). CONCLUSION: Plant use is based on odour, and plant morphological characteristics and is embedded in a complex cultural context based on indigenous Amerindian beliefs. It is suggested that the medicinal plants exerted a physiological action on the hunter or his dog. Some of the plants mentioned contain chemicals that may explain the ethnomedicinal and ethnoveterinary use. For instance some of the plants influence the immune system or are effective against internal and external parasites. Plant baths may contribute to the health and well being of the hunting dogs. [Abstract/Link to Full Text]

Jones BM
Changes in cytokine production in healthy subjects practicing Guolin Qigong : a pilot study.
BMC Complement Altern Med. 2001;18.
BACKGROUND: Guolin Qigong is a combination of meditation, controlled breathing and physical movement designed to control the vital energy (qi) of the body and consequently to improve spiritual, physical and mental health. Practice of Qigong has been reported to alter immunological function, but there have been few studies of its effects on cytokines, the key regulators of immunity. METHODS: Numbers of peripheral blood cytokine-secreting cells were determined by ELISPOT in 19 healthy volunteers aged 27 - 55, before they were taught the practice of Qigong and after 3, 7 and 14 weeks of daily practice. The effect of Qigong on blood cortisol was also examined. RESULTS: Numbers of IL4 and IL12-secreting cells remained stable. IL6 increased at 7 weeks and TNFalpha increased in unstimulated cultures at 3 and 7 weeks but decreased at these times in LPS and SAC-stimulated cultures. Of particular interest, IFNgamma-secreting cells increased and IL10-secreting cells decreased in PHA-stimulated cultures, resulting in significant increases in the IFNgamma:IL10 ratio. Cortisol, a known inhibitor of type 1 cytokine production, was reduced by practicing Qigong. CONCLUSION: These preliminary studies in healthy subjects, although not necessarily representative of a randomized healthy population and not including a separate control group, have indicated that blood levels of the stress-related hormone cortisol may be lowered by short-term practice of Qigong and that there are concomitant changes in numbers of cytokine-secreting cells. Further studies of the effect of Qigong in patients with clinical diseases known to be associated with type 2 cytokine predominance are merited. [Abstract/Link to Full Text]

Donaldson MS, Speight N, Loomis S
Fibromyalgia syndrome improved using a mostly raw vegetarian diet: an observational study.
BMC Complement Altern Med. 2001;17.
BACKGROUND: Fibromyalgia engulfs patients in a downward, reinforcing cycle of unrestorative sleep, chronic pain, fatigue, inactivity, and depression. In this study we tested whether a mostly raw vegetarian diet would significantly improve fibromyalgia symptoms. METHODS: Thirty people participated in a dietary intervention using a mostly raw, pure vegetarian diet. The diet consisted of raw fruits, salads, carrot juice, tubers, grain products, nuts, seeds, and a dehydrated barley grass juice product. Outcomes measured were dietary intake, the fibromyalgia impact questionnaire (FIQ), SF-36 health survey, a quality of life survey (QOLS), and physical performance measurements. RESULTS: Twenty-six subjects returned dietary surveys at 2 months; 20 subjects returned surveys at the beginning, end, and at either 2 or 4 months of intervention; 3 subjects were lost to follow-up. The mean FIQ score (n = 20) was reduced 46% from 51 to 28. Seven of the 8 SF-36 subscales, bodily pain being the exception, showed significant improvement (n = 20, all P for trend < 0.01). The QOLS, scaled from 0 to 7, rose from 3.9 initially to 4.9 at 7 months (n = 20, P for trend 0.000001). Significant improvements (n = 18, P < 0.03, paired t-test) were seen in shoulder pain at rest and after motion, abduction range of motion of shoulder, flexibility, chair test, and 6-minute walk. 19 of 30 subjects were classified as responders, with significant improvement on all measured outcomes, compared to no improvement among non-responders. At 7 months responders' SF-36 scores for all scales except bodily pain were no longer statistically different from norms for women ages 45-54. CONCLUSION: This dietary intervention shows that many fibromyalgia subjects can be helped by a mostly raw vegetarian diet. [Abstract/Link to Full Text]

Linde K, Hondras M, Vickers A, ter Riet G, Melchart D
Systematic reviews of complementary therapies - an annotated bibliography. Part 3: homeopathy.
BMC Complement Altern Med. 2001;14.
BACKGROUND: Complementary therapies are widespread but controversial. We aim to provide a comprehensive collection and a summary of systematic reviews of clinical trials in three major complementary therapies (acupuncture, herbal medicine, homeopathy). This article is dealing with homeopathy. Potentially relevant reviews were searched through the register of the Cochrane Complementary Medicine Field, the Cochrane Library, Medline, and bibliographies of articles and books. To be included articles had to review prospective clinical trials of homeopathy; had to describe review methods explicitly; had to be published; and had to focus on treatment effects. Information on conditions, interventions, methods, results and conclusions was extracted using a pretested form and summarized descriptively. RESULTS: Eighteen out of 22 potentially relevant reviews preselected in the screening process met the inclusion criteria. Six reviews addressed the question whether homeopathy is effective across conditions and interventions. The majority of available trials seem to report positive results but the evidence is not convincing. For isopathic nosodes for allergic conditions, oscillococcinum for influenza-like syndromes and galphimia for pollinosis the evidence is promising while in other areas reviewed the results are equivocal. INTERPRETATION: Reviews on homeopathy often address general questions. While the evidence is promising for some topics the findings of the available reviews are unlikely to end the controversy on this therapy. [Abstract/Link to Full Text]

Linde K, ter Riet G, Hondras M, Vickers A, Saller R, Melchart D
Systematic reviews of complementary therapies - an annotated bibliography. Part 2: herbal medicine.
BMC Complement Altern Med. 2001;15.
BACKGROUND: Complementary therapies are widespread but controversial. We aim to provide a comprehensive collection and a summary of systematic reviews of clinical trials in three major complementary therapies (acupuncture, herbal medicine, homeopathy). This article is dealing with herbal medicine. Potentially relevant reviews were searched through the register of the Cochrane Complementary Medicine Field, the Cochrane Library, Medline, and bibliographies of articles and books. To be included articles had to review prospective clinical trials of herbal medicines; had to describe review methods explicitly; had to be published; and had to focus on treatment effects. Information on conditions, interventions, methods, results and conclusions was extracted using a pre-tested form and summarized descriptively. RESULTS: From a total of 79 potentially relevant reviews pre-selected in the screening process 58 met the inclusion criteria. Thirty of the reports reviewed ginkgo (for dementia, intermittent claudication, tinnitus, and macular degeneration), hypericum (for depression) or garlic preparations (for cardiovascular risk factors and lower limb atherosclerosis). The quality of primary studies was criticized in the majority of the reviews. Most reviews judged the available evidence as promising but definitive conclusions were rarely possible. CONCLUSIONS: Systematic reviews are available on a broad range of herbal preparations prescribed for defined conditions. There is very little evidence on the effectiveness of herbalism as practised by specialist herbalists who combine herbs and use unconventional diagnosis. [Abstract/Link to Full Text]

Recent Articles in Evidence-based Complementary and Alternative Medicine

Vojdani A, Erde J
Regulatory T cells, a potent immunoregulatory target for CAM researchers: the ultimate antagonist (I).
Evid Based Complement Alternat Med. 2006 Mar;3(1):25-30.
Over the past decade, great interest has been given to regulatory T (T(reg)) cells. A vast body of evidence has shown the existence and highlighted the importance of T(reg) cells in the active suppression of immune system responses. This form of immunoregulation is the dominant means utilized by the immune system to reach a harmony between reciprocal response processes in order to ensure adequate host defense with minimal host detriment. Therapeutically targeting T(reg) cells is a direct and powerful means to manipulate the immune system to achieve beneficial effects on various disease pathologies, including allergy, autoimmunity and cancer, as well as the facilitation of organ transplantation. This powerful target for immunoregulation is of much concern to practitioners and researchers of complementary and alternative medicine because it allows a great deal of control and certainty in dealing with the prevalence of debilitating immune system-related disorders for which there has been little remedy outside of Western Medicine. [Abstract/Link to Full Text]

Bellavite P, Conforti A, Pontarollo F, Ortolani R
Immunology and homeopathy. 2. Cells of the immune system and inflammation.
Evid Based Complement Alternat Med. 2006 Mar;3(1):13-24.
Here we describe the results of some experimental laboratory studies aimed at verifying the efficacy of high dilutions of substances and of homeopathic medicines in models of inflammation and immunity. Studies carried out on basophils, lymphocytes, granulocytes and fibroblasts are reviewed. This approach may help to test under controlled conditions the main principles of homeopathy such as 'similarity' of drug action at the cellular level and the effects of dilution/dynamization on the drug activity. The current situation is that few and rather small groups are working on laboratory models for homeopathy. Regarding the interpretation of data in view of the simile principle, we observe that there are different levels of similarity and that the laboratory data give support to this principle, but have not yet yielded the ultimate answer to the action mechanism of homeopathy. Evidence of the biological activity in vitro of highly diluted-dynamized solutions is slowly accumulating, with some conflicting reports. It is our hope that this review of literature unknown to most people will give an original and useful insight into the 'state-of-the-art' of homeopathy, without final conclusions 'for' or 'against' this modality. This kind of uncertainty may be difficult to accept, but is conceivably the most open-minded position now. [Abstract/Link to Full Text]

Chiappelli F, Prolo P, Rosenblum M, Edgerton M, Cajulis OS
Evidence-based research in complementary and alternative medicine II: the process of evidence-based research.
Evid Based Complement Alternat Med. 2006 Mar;3(1):3-12.
It is a common practice in contemporary medicine to follow stringently the scientific method in the process of validating efficacy and effectiveness of new or improved modes of treatment intervention. It follows that these complementary or alternative interventions must be validated by stringent research before they can be reliably integrated into Western medicine. The next decades will witness an increasing number of evidence-based research directed at establishing the best available evidence in complementary and alternative medicine (CAM). This second paper in this lecture series examines the process of evidence-based research (EBR) in the context of CAM. We outline the fundamental principles, process and relevance of EBR, and its implication to CAM. We underscore areas of future development in EBR. We note that the main problem of applying EBR to CAM at present has to do with the fact that the contribution of EBR can be significant only to the extent to which studies used in the process of EBR are of good quality. All too often CAM research is not of sufficient quality to warrant the generation of a consensus statement. EBR, nevertheless, can contribute to CAM by identifying current weaknesses of CAM research. We present a revised instrument to assess quality of the literature. [Abstract/Link to Full Text]

Goldstein MS, Brown ER, Ballard-Barbash R, Morgenstern H, Bastani R, Lee J, Gatto N, Ambs A
The use of complementary and alternative medicine among california adults with and without cancer.
Evid Based Complement Alternat Med. 2005 Dec;2(4):557-65.
This article examines the extent and correlates of complementary and alternative medicine (CAM) use among a population-based sample of California adults that is highly diverse in terms of sociodemographic characteristics and health status. As a follow-up to a state-wide health survey of 55,428 people, 9187 respondents were interviewed by phone regarding their use of 11 different types of CAM providers, special diets, dietary supplements, mind-body interventions, self-prayer and support groups. The sample included all participants in the initial survey who reported a diagnosis of cancer, all the non-white respondents, as well as a random sample of all the white respondents. The relation of CAM use to the respondents' demographic characteristics and health status is assessed. CAM use among Californians is generally high, and the demographic factors associated with high rates of CAM use are the same in California as have been found in other studies. Those reporting a diagnosis of cancer and those who report other chronic health problems indicate a similar level of visits to CAM providers. However, those with cancer are less likely to report using special diets, and more likely to report using support groups and prayer. Health status, gender, ethnicity and education have an independent impact upon CAM use among those who are healthy as well as those who report suffering from chronic health problems, although the precise relation varies by the type of CAM used. [Abstract/Link to Full Text]

Azaizeh H, Ljubuncic P, Portnaya I, Said O, Cogan U, Bomzon A
Fertilization-induced changes in growth parameters and antioxidant activity of medicinal plants used in traditional Arab medicine.
Evid Based Complement Alternat Med. 2005 Dec;2(4):549-56.
In response to increased popularity and greater demand for medicinal plants, a number of conservation groups are recommending that wild medicinal plants be brought into cultivation systems. We collected four medicinal herbs Cichorium pumilum, Eryngium creticum, Pistacia palaestina and Teucrium polium used in traditional Arab medicine for greenhouse cultivation to assess the effects of different fertilization regimes on their growth and antioxidant activity. Wild seedlings were collected and fertilized with either 100% Hoagland solution, 50% Hoagland solution, 20% Hoagland solution or irrigated with tap water. Plant height was measured and the number of green leaves and branches counted weekly. Thereafter, the aboveground parts of plants were harvested for preparing a water-soluble powder extracts of which antioxidant activity was measured by their ability to suppress the oxidation of beta-carotene. Of the fertilization regimes, we found either 20 or 50% Hoagland solution produced the most consistent response of the plant growth parameters. All powders prepared from the four wild growing plants inhibited oxidation of beta-carotene. Increasing the amount of fertilizer caused a significant concentration-dependent increase in antioxidant activity of the cultivated T. polium compared with the wild type. In contrast, increasing the amount of fertilizer caused a significant concentration-dependent reduction in the antioxidant activity of powders prepared from the cultivated E. creticum when compared with wild plants. Our results showed that cultivation success should not rely solely on parameters of growth but should incorporate assessment related to indices of therapeutic potential. [Abstract/Link to Full Text]

Khuda-Bukhsh AR, Pathak S, Guha B, Karmakar SR, Das JK, Banerjee P, Biswas SJ, Mukherjee P, Bhattacharjee N, Choudhury SC, Banerjee A, Bhadra S, Mallick P, Chakrabarti J, Mandal B
Can homeopathic arsenic remedy combat arsenic poisoning in humans exposed to groundwater arsenic contamination?: a preliminary report on first human trial.
Evid Based Complement Alternat Med. 2005 Dec;2(4):537-48.
Groundwater arsenic (As) has affected millions of people globally distributed over 20 countries. In parts of West Bengal (India) and Bangladesh alone, over 100 million people are at risk, but supply of As-free water is grossly inadequate. Attempts to remove As by using orthodox medicines have mostly been unsuccessful. A potentized homeopathic remedy, Arsenicum Album-30, was administered to a group of As affected people and thereafter the As contents in their urine and blood were periodically determined. The activities of various toxicity marker enzymes and compounds in the blood, namely aspartate amino transferase, alanine amino transferase, acid phosphatase, alkaline phosphatase, lipid peroxidation and reduced glutathione, were also periodically monitored up to 3 months. The results are highly encouraging and suggest that the drug can alleviate As poisoning in humans. [Abstract/Link to Full Text]

Han Y, Son SJ, Akhalaia M, Platonov A, Son HJ, Lee KH, Yun YS, Song JY
Modulation of radiation-induced disturbances of antioxidant defense systems by ginsan.
Evid Based Complement Alternat Med. 2005 Dec;2(4):529-36.
There are numerous studies to indicate that irradiation induces reactive oxygen species (ROS), which play an important causative role in radiation damage of the cell. We evaluated the effects of ginsan, a polysaccharide fraction extracted from Panax ginseng, on the gamma-radiation induced alterations of some antioxidant systems in the spleen of Balb/c mice. On the 5th day after sublethal whole-body irradiation, homogenized spleen tissues of the irradiated mice expressed only marginally increased mRNA levels of Mn-SOD (superoxide dimutase) in contrast to Cu/Zn-SOD, however, catalase mRNA was decreased by approximately 50% of the control. In vivo treatment of non-irradiated mice with ginsan (100 mg kg(-1), intraperitoneal administration) had no significant effect, except for glutathione peroxidase (GPx) mRNA, which increased to 144% from the control. However, the combination of irradiation with ginsan effectively increased the SODs and GPx transcription as well as their protein expressions and enzyme activities. In addition, the expression of heme oxygenase-1 and non-protein thiol induced by irradiation was normalized by the treatment of ginsan. Evidence indicated that transforming growth factor-beta and other important cytokines such as IL-1, TNF and IFN-gamma might be involved in evoking the antioxidant enzymes. Therefore, we propose that the modulation of antioxidant enzymes by ginsan was partly responsible for protecting the animal from radiation, and could be applied as a therapeutic remedy for various ROS-related diseases. [Abstract/Link to Full Text]

Tsao JC, Meldrum M, Bursch B, Jacob MC, Kim SC, Zeltzer LK
Treatment expectations for CAM interventions in pediatric chronic pain patients and their parents.
Evid Based Complement Alternat Med. 2005 Dec;2(4):521-7.
Patient expectations regarding complementary and alternative medicine (CAM) interventions have important implications for treatment adherence, attrition and clinical outcome. Little is known, however, about parent and child treatment expectations regarding CAM approaches for pediatric chronic pain problems. The present study examined ratings of the expected benefits of CAM (i.e. hypnosis, massage, acupuncture, yoga and relaxation) and conventional medicine (i.e. medications, surgery) interventions in 45 children (32 girls; mean age = 13.8 years +/- 2.5) and parents (39 mothers) presenting for treatment at a specialty clinic for chronic pediatric pain. Among children, medications and relaxation were expected to be significantly more helpful than the remaining approaches (P < 0.01). However, children expected the three lowest rated interventions, acupuncture, surgery and hypnosis, to be of equal benefit. Results among parents were similar to those found in children but there were fewer significant differences between ratings of the various interventions. Only surgery was expected by parents to be significantly less helpful than the other approaches (P < 0.01). When parent and child perceptions were compared, parents expected hypnosis, acupuncture and yoga, to be more beneficial than did children, whereas children expected surgery to be more helpful than did parents (P < 0.01). Overall, children expected the benefits of CAM to be fairly low with parents' expectations only somewhat more positive. The current findings suggest that educational efforts directed at enhancing treatment expectations regarding CAM, particularly among children with chronic pain, are warranted. [Abstract/Link to Full Text]

Haddad PS, Azar GA, Groom S, Boivin M
Natural health products, modulation of immune function and prevention of chronic diseases.
Evid Based Complement Alternat Med. 2005 Dec;2(4):513-20.
The immune system is increasingly found to be involved in the development of several chronic illnesses, for which allopathic medicine has provided limited tools for treatment and especially prevention. In that context, it appears worthwhile to target the immune system in order to modulate the risk of certain chronic illnesses. Meanwhile, natural health products (NHPs) are generating renewed interest, particularly in the prevention and treatment of several chronic diseases. Over 20 scientists from fields related to immune function and NHPs were thus convened to establish the state of knowledge on these subjects and to explore future research directions. This review summarizes the result of discussions held during the symposium. It thus seeks to be thought provoking rather than to comprehensively cover such broad areas of research. Notably, a brief overview of the immune system is presented, including potentially useful targets and strategies to keep it in an equilibrated state, in order to prevent certain disorders. The pertinence and limitations of targeting the immune system to prevent chronic diseases is also discussed. The paper then discusses the usefulness and limitations of current experimental tools available to study the immune modulating effects of NHPs. Finally, a concise review of some of the most studied NHPs showing promising immunomodulatory activity is given, and avenues for future research are described. [Abstract/Link to Full Text]

Iribarren J, Prolo P, Neagos N, Chiappelli F
Post-traumatic stress disorder: evidence-based research for the third millennium.
Evid Based Complement Alternat Med. 2005 Dec;2(4):503-12.
The stress that results from traumatic events precipitates a spectrum of psycho-emotional and physiopathological outcomes. Post-traumatic stress disorder (PTSD) is a psychiatric disorder that results from the experience or witnessing of traumatic or life-threatening events. PTSD has profound psychobiological correlates, which can impair the person's daily life and be life threatening. In light of current events (e.g. extended combat, terrorism, exposure to certain environmental toxins), a sharp rise in patients with PTSD diagnosis is expected in the next decade. PTSD is a serious public health concern, which compels the search for novel paradigms and theoretical models to deepen the understanding of the condition and to develop new and improved modes of treatment intervention. We review the current knowledge of PTSD and introduce the role of allostasis as a new perspective in fundamental PTSD research. We discuss the domain of evidence-based research in medicine, particularly in the context of complementary medical intervention for patients with PTSD. We present arguments in support of the notion that the future of clinical and translational research in PTSD lies in the systematic evaluation of the research evidence in treatment intervention in order to insure the most effective and efficacious treatment for the benefit of the patient. [Abstract/Link to Full Text]

Capodice JL, Bemis DL, Buttyan R, Kaplan SA, Katz AE
Complementary and alternative medicine for chronic prostatitis/chronic pelvic pain syndrome.
Evid Based Complement Alternat Med. 2005 Dec;2(4):495-501.
To discuss challenges concerning treatment for chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) and review complementary and alternative medical (CAM) therapies being evaluated for this condition, we performed a comprehensive search of articles published from 1990-2005 using the PubMed, Medline databases. Data from the articles were abstracted and pooled by subject. Keywords cross-searched with CP/CPPS included: complementary, alternative, integrative, therapies, interventions, nutrition, antioxidants, herbs, supplements, biofeedback and acupuncture. Listed articles with no abstracts were not included. Various CAM therapies for CP/CPPS exist including biofeedback, acupuncture, hyperthermia and electrostimulation. Additionally, a variety of in vitro and in vivo studies testing herbal and nutritional supplements were found. Saw palmetto, cernilton and quercetin were the most frequently tested supplements for CP/CPPS. Although many CAM therapies demonstrate positive preliminary observations as prospective treatments for CP/CPPS, further exploratory studies including more randomized, controlled trials are necessary for significant validation as treatment options for this complex disorder. [Abstract/Link to Full Text]

Chiappelli F
The molecular immunology of mucositis: implications for evidence-based research in alternative and complementary palliative treatments.
Evid Based Complement Alternat Med. 2005 Dec;2(4):489-94.
The terms 'mucositis' and 'stomatitis' are often used interchangeably. Mucositis, however, pertains to pharyngeal-esophago-gastrointestinal inflammation that manifests as red, burn-like sores or ulcerations throughout the mouth. Stomatitis is an inflammation of the oral tissues proper, which can present with or without sores, and is made worse by poor dental hygiene. Mucositis is observed in a variety of immunosuppressed patients, but is most often consequential to cancer therapy. It appears as early as the third day of intervention, and is usually established by Day 7 of treatment. Mucositis increases mortality and morbidity and contributes to rising health care costs. The precise immune components involved in the etiology of mucositis are unclear, but evidence-based research (EBR) data has shown that applications of granulocyte-macrophage-colony stimulating factor prevent the onset or the exacerbation of oropharyngeal mucositis. The molecular implications of this observation are discussed from the perspective of future developments of complementary and alternative treatments for this condition. It must be emphasized that this article is meant to be neither a review on mucositis and the various treatments for it, nor a discussion paper on its underlying molecular immunology. It is a statement of the implications of EBR for CAM-based interventions for mucositis. It explores and discusses the specific domain of molecular immunology in the context of mucositis and its direct implications for EBR research in CAM-based treatments for mucositis. [Abstract/Link to Full Text]

Azeemi ST, Raza SM
A critical analysis of chromotherapy and its scientific evolution.
Evid Based Complement Alternat Med. 2005 Dec;2(4):481-8.
Chromotherapy is a method of treatment that uses the visible spectrum (colors) of electromagnetic radiation to cure diseases. It is a centuries-old concept used successfully over the years to cure various diseases. We have undertaken a critical analysis of chromotherapy and documented its scientific evolution to date. A few researchers have tried to discover the underlying scientific principles, but without quantitative study. Sufficient published material can be found about the subject that provides a complete system of treatment focused on the treatment methodologies and healing characteristics of colors. A number of studies have elaborated the relationship between the human body and colors. We also show the possibility of carrying out diverse research into chromotherapy that is pertinent to deciphering the quantum mechanical dipole moment of water molecules. The quantum mechanical dipole moment as a result of the absorption of different colors, we conjecture, produces charge quantization phenomena. This review illustrates that the development of science in the field of electromagnetic radiation/energy can be very helpful in discovering new dimensions of this old theory. [Abstract/Link to Full Text]

Saad B, Azaizeh H, Said O
Tradition and perspectives of arab herbal medicine: a review.
Evid Based Complement Alternat Med. 2005 Dec;2(4):475-9.
Complementary and Alternative Medicine (CAM), including herbal medicine, are popular in the general population worldwide. Parallel to the increasing interest in 'modern' CAM therapies and the historical importance of Arab medicine, there is also a similar trend in research activities dealing with the efficacy and safety of medicinal plants in our region. Historical and current studies and surveys indicate that the Eastern region of the Mediterranean has been distinguished throughout the generations with a rich inventory of natural medicinal herbs. It is well documented that indigenous Arab medicine has contributed greatly to the development of modern medicine in Europe and remains one of the closest forms of original European medicine. The rapid increase in consumption of herbal remedies worldwide has been stimulated by several factors, including the notion that all herbal products are safe and effective. This article presents a systematic review on traditional Arab medicine including historical background, medical innovations introduced by Arab physicians in the field of safety and efficacy of herbal medicine and a state-of-the-art description of traditional Arab herbal medicine in the Mediterranean region. [Abstract/Link to Full Text]

Patwardhan B, Warude D, Pushpangadan P, Bhatt N
Ayurveda and traditional Chinese medicine: a comparative overview.
Evid Based Complement Alternat Med. 2005 Dec;2(4):465-73.
Ayurveda, the traditional Indian medicine (TIM) and traditional Chinese medicine (TCM) remain the most ancient yet living traditions. There has been increased global interest in traditional medicine. Efforts to monitor and regulate herbal drugs and traditional medicine are underway. China has been successful in promoting its therapies with more research and science-based approach, while Ayurveda still needs more extensive scientific research and evidence base. This review gives an overview of basic principles and commonalities of TIM and TCM and discusses key determinants of success, which these great traditions need to address to compete in global markets. [Abstract/Link to Full Text]

Adams JD, Garcia C
Spirit, mind and body in Chumash healing.
Evid Based Complement Alternat Med. 2005 Dec;2(4):459-63.
This article discusses the importance of the spirit and mind in health and well-being among Chumash people. Prayer was the first step in healing since prayer invites the participation of God. Initiation practices are discussed that encouraged young people to develop the maturity and spiritual strength to become productive members of society. Pictographs were used in healing usually not only as a relaxation therapy, but also as a mode of education. A supportive environment was an important factor in Chumash health care, since the support of friends helps, comforts and relieves anxiety that is detrimental to healing. [Abstract/Link to Full Text]

Chiappelli F, Prolo P, Cajulis OS
Evidence-based research in complementary and alternative medicine I: history.
Evid Based Complement Alternat Med. 2005 Dec;2(4):453-8.
Contemporary Western medicine has witnessed a fragmentation of our conceptualization of the medical endeavor into 'traditional medicine' and 'non-traditional medicine'. The former is meant to refer to the Western medical tradition, the latter encompasses both 'complementary' and 'alternative' medical practices. Complementary medicine complements conventional medical treatments, and alternative modes of medical interventions are meant to replace traditional Western medicine. Evidence-based research must be directed at establishing the best available evidence in complementary and alternative medicine. This paper is the first of a set of four 'lectures' that reviews the process of evidence-based research, and discusses its implications and applications for the early decades of the 21st century. The purpose of this paper is to introduce the series by examining some of the historical and philosophical foundations of this research endeavor. [Abstract/Link to Full Text]

Bellavite P, Conforti A, Piasere V, Ortolani R
Immunology and homeopathy. 1. Historical background.
Evid Based Complement Alternat Med. 2005 Dec;2(4):441-52.
Homeopathy was born as an experimental discipline, as can be seen from the enormous amount of homeopathic data collected over more than two centuries. However, the medical tradition of homeopathy has been separated from that of conventional science for a long time. Conventional scientific wisdom dictates that homeopathy should have no effect above placebo but experiments on ultra-high dilutions of solutes together with some clinical data suggest the intriguing possibility that it might do in some circumstances. Today, an osmotic process between disciplines, previously seen as in conflict, is facilitated because over the last few decades homeopathy has initiated the methods of current medical science and a substantial number of experimental studies-at molecular, cellular and clinical levels-are available. One area of dialogue and of common progress is that of inflammation and immunity, probably because these are closely related to the traditional 'vital force' of the body's self-healing power. In a series of papers we review the historical origins of homeopathy, the laboratory and animal models related to the field of immunopharmacology, the clinical evidence in favor and against the use of homeopathy in the inflammatory diseases and the hypotheses regarding its action mechanism(s). Finally, we will enlighten the specific characteristics of the homeopathic approach, which places great emphasis on identifying a cure for the whole organism. [Abstract/Link to Full Text]

Olalde Rangel JA, Magarici M, Amendola F, del Castillo O
The Systemic Theory of Living Systems. Part IV: Systemic Medicine--The Praxis.
Evid Based Complement Alternat Med. 2005 Dec;2(4):429-39.
This fourth lecture illustrates the praxis and results of Systemic Medicine (SM) in various therapeutic applications. SM's success has made it popular throughout Venezuela and Puerto Rico. The treatment of over 300,000 patients by 150 orthodox MD's, trained and qualified in SM, in 35 medical establishments with above average results corroborate its effectiveness as an eCAM in chronic degenerative diseases. Herein we provide a synopsis of results obtained in four such pathologies-the journal's necessary space restrictions somewhat limiting content-as well as clinical and photographic evidence. The validity of any medical theory is substantiated by its degree of effectivity and success. The workability of evidence-based SM corroborates Systemic Theory's transcendence. [Abstract/Link to Full Text]

Cooper EL
ECAM is waiting for eCAM.
Evid Based Complement Alternat Med. 2005 Dec;2(4):427-8. [Abstract/Link to Full Text]

Lewith G
Complementary medicine research unit.
Evid Based Complement Alternat Med. 2005 Sep;2(3):399-407. [Abstract/Link to Full Text]

Ghassemi J
Finding the evidence in CAM: a student's perspective.
Evid Based Complement Alternat Med. 2005 Sep;2(3):395-7.
This commentary offers a future health care provider's perspective on the role of complementary and alternative medicine (CAM) in Western (namely, in US) medical education and practice. As a student of both public health and medicine in the United States, Jeffrey Ghassemi is interested in CAM's contribution to improving medical practice and teaching. The commentary highlights the ambiguous definitions of CAM to Westerners despite the rising popularity of and expenditures for alternative modalities of care. It then argues for collaboration between alternative and established medical communities to ascertain the scientific merits of CAM. It concludes by calling for a new medical paradigm that embraces the philosophies of both communities to advance education and patient care. [Abstract/Link to Full Text]

Ohnishi ST, Ohnishi T, Nishino K, Tsurusaki Y, Yamaguchi M
Growth inhibition of cultured human liver carcinoma cells by Ki-energy (life-energy): scientific evidence for Ki-effects on cancer cells.
Evid Based Complement Alternat Med. 2005 Sep;2(3):387-93.
'Ki-energy' (life-energy) is believed to increase the immune activity of its practitioners. It has also been shown to cause neuropsychological effects. We undertook this study to obtain objective and scientific evidence as to whether or not a 'Ki-effect' could inhibit the growth of cultured cancer cells. Cultured human liver carcinoma cells, HepG2, were used. A Japanese Ki-expert held his fingers toward the cells in culture dishes for 5 or 10 min. After culturing for 24 h, we measured cell numbers, protein concentration per cell, certain mRNA expressions and the synthesis of regucalcin. The results were compared with those for control cells (non-treated cells). We found that the number of cells in the Ki-exposed groups were less than those in the controls by 30.3 and 40.6% with 5 and 10 min Ki-exposure, respectively. The protein content per cell in the Ki-exposed groups (5 and 10 min) was higher than that in the control groups by 38.8 and 62.9%, respectively. These results were statistically significant. Using RT-PCR, we found that the mRNA expression for c-myc, a tumor stimulator gene, was decreased, while that for regucalcin, which suppresses DNA synthesis, was increased. Our molecular biological studies and mathematical model analysis demonstrated that Ki-energy inhibited cancer cell division. The data also indicate that the Ki-effects involve some form of infrared radiation from the human body. This study suggests the possibility that Ki-energy may be beneficial for cancer patients because it suppresses cancer cell growth, and at the same time, it stimulates immune functions of the patients. [Abstract/Link to Full Text]

Karimi G, Ramezani M, Tahoonian Z
Cisplatin nephrotoxicity and protection by milk thistle extract in rats.
Evid Based Complement Alternat Med. 2005 Sep;2(3):383-6.
The protective effect of methanolic extract of milk thistle seeds and silymarin against cisplatin-induced renal toxicity in male rats after a single intraperitoneal injection of 3 mg kg cisplatin were studied. Over 5 days, cisplatin-treated rats showed tubular necrosis and elevation in blood urea nitrogen (BUN) and serum creatinine (Scr). Pretreatment of animals with silymarin (50 mg kg) or extract (0.6 g kg) 2 h before cisplatin prevented the tubular damage. Rats treated with silymarin or extract 2 h after cisplatin had BUN and Scr significantly lower than those receiving cisplatin, but mild to moderate necrosis was observed. These results suggested that milk thistle may protect against cisplatin-induced renal toxicity and might serve as a novel combination agent with cisplatin to limit renal injury. [Abstract/Link to Full Text]

Punitha IS, Rajendran K, Shirwaikar A, Shirwaikar A
Alcoholic stem extract of Coscinium fenestratum regulates carbohydrate metabolism and improves antioxidant status in streptozotocin-nicotinamide induced diabetic rats.
Evid Based Complement Alternat Med. 2005 Sep;2(3):375-81.
Alcoholic extract of the stems of Coscinium fenestratum, a medicinal plant indigenous to India and Sri Lanka used in ayurveda and siddha medicine for treating diabetes, was studied for its carbohydrate metabolism effect and antioxidant status in streptozotocin-nicotinamide induced type 2 diabetic rats. Oral administration of C. fenestratum stem extract in graded doses caused a significant increase in enzymatic antioxidants such as catalase, superoxide dismutase, glutathione synthetase, peroxidase, and glutathione peroxidase and in the nonenzymatic antioxidants ascorbic acid, ceruloplasmin and tocopherol. Effects of alcoholic extract on glycolytic enzymes such as glucose-6-phosphate dehydrogenase, lactate dehydrogenase and hexokinase showed a significant increase in their levels, whereas a significant decrease was observed in the levels of gluconeogenic enzyme, glucose-6-phosphatase and alanine aminotransferase in treated diabetic rats. Serum creatinine and urea levels also declined significantly. This investigation demonstrates significant antidiabetic activity of C. fenestratum. [Abstract/Link to Full Text]

Inagaki N, Shibata T, Itoh T, Suzuki T, Tanaka H, Nakamura T, Akiyama Y, Kawagishi H, Nagai H
Inhibition of IgE-dependent mouse triphasic cutaneous reaction by a boiling water fraction separated from mycelium of Phellinus linteus.
Evid Based Complement Alternat Med. 2005 Sep;2(3):369-74.
Phellinus linteus, a mushroom, contains constituents that exhibit potent antitumor effects through activating immune cells. Recently, anti-inflammatory and anti-allergic properties of P. linteus extracts have also been implicated. In the present study, therefore, we separated the constituents of mycelium of P. linteus into five fractions-chloroform-soluble (CF), ethyl acetate-soluble (EA), methanol-soluble (AE), water-soluble (WA) and boiling water-soluble (BW) fractions-and examined their suppressive effects on the IgE-dependent mouse triphasic cutaneous reaction. The triphasic reaction was induced in the ear of BALB/c mice passively sensitized with anti-dinitrophenol IgE by painting with 2,4-dinitrofluorobenzene 24 h later. Ear swelling appeared triphasically with peak responses at 1 h, 24 h and 8 days after the challenge. ME, WA and BW given orally at a dose of 100 mg kg significantly inhibited the first and second phase ear swelling, and BW also inhibited the third phase response. CF only inhibited the second phase. The inhibition by BW was the most potent and almost dose-dependent at doses of 30-300 mg kg. BW also inhibited vascular permeability increase caused by passive cutaneous anaphylaxis and histamine, and ear swelling caused by tumor necrosis factor-alpha. In contrast, BW apparently potentiated the production of interleukin-4 and interferon-gamma from anti-CD3-stimulated mouse splenocytes. These results indicate that BW derived from mycelium of P. linteus contains some constituents with anti-allergic as well as immunopotentiating properties. [Abstract/Link to Full Text]

Cao Y, Shi Y, Zheng Y, Shi M, Lo SK
Blood-nourishing and hard-softening capsule costs less in the management of osteoarthritic knee pain: a randomized controlled trial.
Evid Based Complement Alternat Med. 2005 Sep;2(3):363-8.
The blood-nourishing and hard-softening (BNHS) capsule is a traditional Chinese formula used in the symptomatic treatment of inflammation and pain. We conducted this randomized controlled trial to compare the efficacy of BNHS with other commonly prescribed drugs. We recruited 120 patients from two teaching hospitals; 30 patients in each hospital were randomly assigned to receive BNHS. In one hospital, the 30 controls were given another traditional Chinese drug; whereas a Western medicine (chondroprotection drug/Viartril-s) was used as the control in the other hospital. Intervention was carried out over a period of 4 weeks. Primary outcome measures included self-reported pain level, and changes in stiffness and functional ability as measured by the Western Ontario McMaster Universities Osteoarthritis (WOMAC) index. Mixed models were used for statistical analysis. Substantial improvements in disease-specific symptoms were observed, after 4 weeks of treatment, in patients taking BNHS capsules. As assessed by the WOMAC index, pain level of the BNHS group decreased by 57% [95% confidence interval (CI) = 50, 63], stiffness by 63% (95% CI = 55, 71) and functional ability increased by 56% (95% CI = 50, 63). No significant differences were found in any of the outcome measures between the BNHS group and either of the comparison groups. No severe adverse effects were reported. However, this study lacked a placebo group; therefore, we conclude that BNHS appears to be as effective as commonly prescribed medicines for the relief of pain and dysfunction in knee osteoarthritis patients, but costs a lot less than other Western and herbal drugs in the study. [Abstract/Link to Full Text]

Xie F, Wu CF, Lai WP, Yang XJ, Cheung PY, Yao XS, Leung PC, Wong MS
The osteoprotective effect of Herba epimedii (HEP) extract in vivo and in vitro.
Evid Based Complement Alternat Med. 2005 Sep;2(3):353-61.
Herba epimedii (HEP) is one of the most frequently used herbs prescribed for treatment of osteoporosis in China. In the present study, the in vivo effects of HEP extract on bone metabolism were evaluated using 4-month-old ovariectomized (OVX) or sham-operated (Sham) female Sprague-Dawley rats orally administered with HEP extract (110 mg kgd), 17ss-estrogen (2 mg kgd) or its vehicle for 3 months. HEP extract significantly decreased urinary calcium excretion, suppressed serum alkaline phosphatase (ALP) activity and urinary deoxypyridinoline levels in OVX rats (P < 0.05 versus vehicle-treated OVX rats). Histomorphometric analysis indicated that HEP extract could prevent OVX-induced bone loss by increasing tibial trabecular bone area and decreasing trabecular separation in OVX rats (P < 0.05 versus vehicle-treated OVX group). The in vitro effects of HEP extract were also studied using rat osteoblast-like UMR 106 cells. HEP extract significantly stimulated cell proliferation in a dose-dependent manner (P < 0.01 versus vehicle-treated) and increased ALP activity at 200 microgml (P < 0.01 versus vehicle-treated) in UMR 106 cells. It modulated osteoclastogenesis by increasing osteoprotegrin (OPG) mRNA and decreasing receptor activator of NF-kappaB ligand (RANKL) mRNA expression, resulting in a dose-dependent increase in OPG/RANKL mRNA ratio (P < 0.01 versus vehicle-treated). Taken together, HEP treatment can effectively suppress the OVX-induced increase in bone turnover possibly by both an increase in osteoblastic activities and a decrease in osteoclastogenesis. The present study provides the evidence that HEP can be considered as a complementary and alternative medicine for treatment of post-menopausal osteoporosis. [Abstract/Link to Full Text]

Kim YS, Jun H, Chae Y, Park HJ, Kim BH, Chang IM, Kang SK, Lee HJ
The practice of Korean medicine: an overview of clinical trials in acupuncture.
Evid Based Complement Alternat Med. 2005 Sep;2(3):325-52.
Acupuncture, one of the Oriental medical therapeutic techniques that can be traced back at least 2500 years, is growing in popularity all over the world. Korea has continued to develop its own unique tradition of medicine throughout its long history, and has formed different types of acupuncture methods. The purpose of this review is to summarize clinical case studies in acupuncture and related therapies, such as acupressure, electric acupuncture, auricular acupuncture and moxibustion in Korea. A survey of Korean journals revealed that a total of 124 studies were published from 1983 to 2001. Results obtained from the survey showed that most clinical studies using acupuncture, electric acupuncture, moxibustion and other traditional therapies could alleviate a relatively broad range of medical problems. However, it should be emphasized that almost all clinical case studies published in various local journals did not follow the 'good clinical practice' with respect to regulatory aspects. Since they were not conducted using the randomized double-blinded controls with a large sample size, all the results should be considered as therapeutic indications. This review is an attempt to show the scope of acupuncture in our country and the kind of diseases, after many years of clinical experience, that were deemed valid targets for clinical trials. [Abstract/Link to Full Text]

Chinnock P, Siegfried N, Clarke M
Is evidence-based medicine relevant to the developing world?: Systematic reviews have yet to achieve their potential as a resource for practitioners in developing countries.
Evid Based Complement Alternat Med. 2005 Sep;2(3):321-4. [Abstract/Link to Full Text]

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Ruiz L, Reyes N, Aroche K, Tolosa V, Simanca V, Rogríguez T, Hardy E
Influence of packaging material on the liquid stability of interferon-alpha2b.
J Pharm Pharm Sci. 2005;8(2):207-16.
PURPOSE: In this article we studied the effect of the packaging material on the liquid stability of interferon alpha 2b (rhIFN-alpha2b). METHODS: The compatibility of this cytokine with type I borosilicate glass ampoules was evaluated by ELISA and RP-HPLC, at 4 degrees C and after heat sealing. Additionally, the influence of protein concentration (3 and 10 MIU/ml), buffer species (sodium phosphate, sodium citrate and sodium citrate-phosphate) and additives (polysorbate 80 and EDTA Na(2) x 2H(2)O) were studied in samples with and without contact with chlorobutyl stoppers by RP-HPLC. RESULTS: The compatibility of this cytokine in sodium phosphate buffer, with type I borosilicate glass ampoules showed a significant adsorption at the lowest concentration. This influence was eliminated with a polysorbate 80/benzyl alcohol-based vehicle. The effect of the heat sealing of ampoules on the stability of rhIFN-alpha2b showed two degradation peaks when a volume of 1 ml was dispensed. However, with a lower (0.5 ml) volume, the degradation was not detected. On the other hand, samples in contact with chlorobutyl stoppers increased the apparent degradation rate constant in the range of 6.74 +/- 0.38 to 46.34 +/- 3.11 x 10(3) day-(1). This effect significantly decreased in about 1.2- and 1.1-fold when sodium citrate or sodium citrate-phosphate buffers, respectively, were evaluated. Results from the evaluation of EDTA Na(2) x 2H(2)O or polysorbate 80 showed a similar behavior. These additives reduced the apparent degradation rate constant in the range of 2.01 +/- 0.14 to 25.51 +/- 3.57 x 10(3) day-(1). CONCLUSIONS: The adsorption of the cytokine to type I borosilicate glass ampoules was eliminated with a polysorbate 80/benzyl alcohol-based vehicle, and the deleterious effect of the heat sealing decreased with a lower (0.5 ml) volume. Experimental data indicated that the contact with chlorobutyl stoppers accelerates the degradation of rhIFN-alpha2b. However, protein concentration, buffer species and pharmaceutical excipients can modulate this effect. [Abstract/Link to Full Text]

Zakaria ZA, Sulaiman MR, Somchit MN, Jais AM, Ali DI
The effects of L-arginine, D-arginine, L-name and methylene blue on channa striatus-induced peripheral antinociception in mice.
J Pharm Pharm Sci. 2005;8(2):199-206.
PURPOSE: To determine the involvement of nitric oxide/cyclic guanosine monophosphate (NO/cGMP) pathway in aqueous supernatant of haruan (Channa striatus) fillet (ASH) antinociception using the acetic acid-induced abdominal constriction test. METHODS: The ASH was prepared by soaking fresh haruan fillet in chloroform:methanol (CM) (2/1 (v/v)) for 72 h followed by evaporation of the upper layer supernatant to remove any solvent residues. The supernatant was then subjected to a freeze-drying process (48 h) followed by doses preparation. RESULTS: Subcutaneous (SC) administration of ASH alone (0.170, 0.426 and 1.704 mg/kg) exhibited a dose-dependent antinociception. On the other hand, 20 mg/kg (SC) of L-arginine and MB exhibited a significant nociception and antinociception, while D-arginine and L-NAME did not produce any effect at all. Pre-treatment with L-arginine was found to significantly reverse the three respective doses of ASH antinociception; pre-treatment with D-arginine did not produce any significant change in the ASH activity; pre-treatment with L-NAME only significantly increased the 0.170 and 0.426 mg/kg ASH antinociception; and pre-treatment with MB significantly enhanced the respective doses of ASH antinociception, respectively. Furthermore, co-treatment with L-NAME significantly enhanced the L-arginine reversal effect on 0.426 mg/kg ASH antinociception. In addition, MB significantly reversed the L-arginine nociception on 0.426 mg/kg ASH. CONCLUSIONS: These finding suggest ASH antinociception involves the nitric oxide (NO)/cyclic guanosine monophosphate (cGMP) pathway. The presence of NO was found to reverse ASH antinociceptive activity while blocking of cGMP system enhanced it. [Abstract/Link to Full Text]

Shafaati A, Lucy C
Application of capillary zone electrophoresis with indirect UV detection to the determination of a model drug, vigabatrin, in dosage forms.
J Pharm Pharm Sci. 2005;8(2):190-8.
PURPOSE: Vigabatrin, an anti-epileptic drug with poor UV absorptivity, is used as a model drug to investigate parameters affecting quantitative determination of cationic drugs using capillary zone electrophoresis with indirect UV detection. METHODS: All experiments were performed on a HP 3D CE instrument equipped with an on-column diode array UV absorbance detector. Untreated fused silica capillaries with an inner diameter of 50 microm, an outer diameter of 365 microm, and a total length of 50 cm (41.5 cm to the detector) were used. Indirect UV detection was performed at 214 nm. Operational parameters such as buffer type and concentration, its pH, probe type and concentration, and the impact of co-ions on the efficiency of separation, were studied. Sabrilex sachets and Sabril tablets were subjected to analysis in this study. RESULTS: Optimal separation and quantification of vigabatrin was obtained using 5 mM sodium phosphate buffer containing 5 mM benzyl tri-ethyl ammonium hydroxide (BTEA) at pH 2.2, with 8-aminocaprylic acid as the internal standard. The method was linear over a range of 5-150 microg/ml (r = 0.9911) and a wider range of 100-600 microg/ml (r = 0.9937) concentration of the drug. The relative standard deviation (RSD) of migration time for 10 consecutive injections of a standard solution of vigabatrin was 0.19%. The limit of quantification (LOQ) was 5 microg/ml. CONCLUSIONS: The method was demonstrated for quantification of vigabatrin in both tablet and sachet dosage forms and proved to be a very specific and fast (8 min) means of routine analysis of the drug in dosage forms, in assay or dissolution testing. [Abstract/Link to Full Text]

Goel RK, Singh A, Naidu PS, Mahajan MP, Kulkarni SK
PASS assisted search and evaluation of some azetidin-2-ones as C.N.S. active agents.
J Pharm Pharm Sci. 2005;8(2):182-9.
PURPOSE: The present study evaluates some azetidin-2-ones derivatives for their central nervous system (CNS) modulating activities. The compounds were chosen from a series (5a-o) which were previously synthesized and evaluated for hypolipidemic and antihyperglycemic activity based on the predictions made by the computer software "Prediction of Activity Spectra for Substances (PASS)". MATERIAL AND METHODS: The test compounds were predicted to have a variety of biological activities but those with the best potential for CNS modulating activity were selected for evaluation of a particular CNS activity as 5a for anti-anxiety, 5b, 5n and 5j for nootropic activity and compound 5c anti-catatonic and anti-dyskinetic activities. Test compound 5a was evaluated for anti-anxiety activity in mirrored chamber model and for pentobarbitone induced sleep potentiation in mice. Test compounds 5b, 5n and 5j were evaluated for nootropic activity in mice by examining the effect on transfer latency on elevated plus maze (EPM) in mice and compound 5c was tested for anti-catatonic activity in perphenazine-induced catatonia and anti-dyskinetic effects in reserpine induced orofacial dyskinesia in rats, respectively. RESULTS AND DISCUSSION: The test compound 5a showed significant anxiolytic activity in the mirror chamber paradigm and showed potentiation of the pentobarbitone-induced hypnosis, which was comparable to diazepam. The nootropic activity of compounds 5b, 5n and 5j were found to be significant in elevated and maze test. The test compound 5c significantly prevented the perphenazine-induced catalepsy in a dose dependent manner. Potentiation of anti-catatonic effect of sub-effective dose of L-dopa and reversal of sulpiride-induced catalepsy was also observed by compound 5c. It indicated that the test compound might be showing anticatatonic effect by dopaminergic stimulation probably through D2 dopaminergic receptors. Compound 5c significantly reduced the vacuous chewing movements, tongue protrusions and jaw tremors induced by reserpine. It further supports the dopaminergic agonism by the test compound as reserpine induces oral dyskinetic features by depleting catecholamine (dopamine and nor- epinephrine). CONCLUSION: It was concluded that azetidinones possess considerable CNS activities and can be further explored to find additional CNS active compounds. [Abstract/Link to Full Text]

Dastmalchi S, Garjani A, Maleki N, Sheikhee G, Baghchevan V, Jafari-Azad P, Valizadeh H, Barzegar-Jalali M
Enhancing dissolution, serum concentrations and hypoglycemic effect of glibenclamide using solvent deposition technique.
J Pharm Pharm Sci. 2005;8(2):175-81.
PURPOSE: Glibenclamide is practically insoluble in water and its GI absorption is limited by its dissolution rate. Therefore, to enhance the drug dissolution, serum concentrations and its hypoglycemic effects, it was formulated as solid dispersions and evaluated the relevant in vitro and in vivo parameters. METHODS: The drug solid dispersions were prepared by solvent deposition technique using microcrystalline cellulose as the carrier in different ratios and their dissolution rates were compared to those of pure drug and its physical mixture with carrier. Drug serum concentrations and hypoglycemic effects in rabbits of pure drug, a physical mixture and the corresponding solid dispersion were investigated. In order to elucidate the observed in vitro and in vivo differences, IR spectroscopy and x-ray diffraction patterns of the formulations were studied. RESULTS: The solid dispersion with the drug to carrier ratio of 1:19 showed the highest dissolution rate with the dissolution efficiency (DE) of 44.42 in comparison to pure drug (DE = 3.82), physical mixture (DE = 4.91) and other solid dispersions (DE between 13.85-39.94) and also produced higher drug serum concentrations (more than 4 times at 6th hour post dose) as well as enhanced hypoglycemic effects relative to pure drug and its corresponding physical mixture. CONCLUSIONS: Solvent deposition technique was proved an effective tool of increasing dissolution probably due to enhanced wettability and reduced drug particle size, which in turn led to enhance drug serum concentrations and its hypoglycemic effects. Strong quantitative correlations were established between dissolution parameter and parameters related to serum concentrations as well as hypoglycemic effects. [Abstract/Link to Full Text]

Bertholet P, Gueders M, Dive G, Albert A, Barillaro V, Perly B, Cataldo D, Piel G, Delattre L, Evrard B
The effect of cyclodextrins on the aqueous solubility of a new MMP inhibitor: phase solubility, 1 H-NMR spectroscopy and molecular modeling studies, preparation and stability study of nebulizable solutions.
J Pharm Pharm Sci. 2005;8(2):163-74.
PURPOSE: Ro 28-2653 (RO) is a synthetic inhibitor of matrix metalloproteinases (MMPs), which is potentially effective against bronchial remodeling. Given that this molecule has very poor aqueous solubility, different cyclodextrins (CDs) have been tested to increase its solubility. The aim of this study was to prepare and to characterize inclusion complexes between RO and CDs, in order to develop nebulizable solutions. METHODS: The complex formation was investigated by phase solubility studies. (1)H-NMR spectroscopy and molecular modeling studies were carried out to elucidate the structure of the inclusion complex between RO and dimethyl-beta-CD (DIMEB). Nebulizable solutions of RO were developed with CDs and a stability study was performed over 9 months. RESULTS: The phase solubility studies showed that beta-CD and its derivatives form a 1:2 complex with RO, whereas gamma-CD includes RO with a 1:1 stoichiometry and a weak stability constant. T-ROESY spectra showed that DIMEB is able to complex two RO substituents (nitrophenyl and biphenyl groups) with preferential orientations, while molecular modeling demonstrated that the configurations observed with (1)H-NMR are energetically favorable, especially owing to H-bond formation between RO and DIMEB. Two CDs were selected to develop nebulizable solutions of RO and the stability study demonstrated that RO degradation in solution is strongly dependent on the concentration of the 1:2 inclusion complex. CONCLUSIONS: CDs are able to include RO and to improve its aqueous solubility. The beta-CD derivatives can be used to formulate nebulizable solutions of RO, the stability of which depends on the concentration of the 1:2 complex. [Abstract/Link to Full Text]

Rangel-Yagui CO, Pessoa A, Tavares LC
Micellar solubilization of drugs.
J Pharm Pharm Sci. 2005;8(2):147-65.
PURPOSE: Micellar solubilization is a powerful alternative for dissolving hydrophobic drugs in aqueous environments. In this work, we provide an insight into this subject. METHODS: A concise review of surfactants and micelles applications in pharmacy was carried out. RESULTS: Initially, a description of surfactants and aqueous micellar systems is presented. Following, an extensive review on micellar drug solubilization, including both the principles involved on this phenomenon and the work already done regarding solubilization of drugs by micelles is presented. The application of micelles in drug delivery, in order to minimize drug degradation and loss, to prevent harmful side effects, and to increase drug bioavailability, is also presented. Special emphasis is given to the more recent use of polymeric micelles. Finally, we briefly discuss the importance of surfactants and micelles as biological systems models as well as its application in micellar catalysis. CONCLUSIONS: As can be seen from the review presented, the use of micelles in pharmacy is an important tool that finds numerous applications. [Abstract/Link to Full Text]

Yusuff K, Awotunde M
The frequency of drug history documentation in an institutionalized tertiary care setting in Nigeria.
J Pharm Pharm Sci. 2005;8(2):141-6.
PURPOSE: The study set out to investigate the frequency of institutionalized patients' drug history documentation in a tertiary care setting in Nigeria and identify opportunities for intervention to improve documentation. METHOD: A cross-sectional retrospective study was carried on June 1st to August 31st 2002 at a 900-bed tertiary care facility located in South Western Nigeria. Stratified random samples of 450 case notes of institutionalized patients who were admitted, discharged or who died at the study site was evaluated for comprehensiveness of drug history documentation with the aid of two pre-piloted data collection forms. RESULT: Drug history documentation was done mainly by attending physicians in all 450 case notes studied (100%). Past use of prescription, over-the-counter and herbal drugs were documented in 33.3%, 12.9% and 6.9% of patients respectively. The dose, frequency and duration of use were documented in 6.4% and 8.4% while past side effects experienced were documented in only 1.6%. Allergy to drug(s), food and chemical(s) were documented in 1.4%, 1.8% and 0.8% respectively. Documentation of use of alcohol, cigarette and illicit drugs were done in 36.6%, 23.2% and 4.2% of patients. Patient adherence with drugs used in the past and source(s) of purchase of these drugs were documented in only 10.2% and 6.6% of patients respectively. CONCLUSION: The documentation of institutionalized patients' drug history in Nigeria is currently not as detailed as it should be. A planned intervention is on going to identify factors responsible for the observed inadequacy and assess the impact of pharmacists' involvement on the quality of drug history documentation. [Abstract/Link to Full Text]

Ochoa L, Igartua M, Hernández RM, Gascón AR, Pedraz JL
Preparation of sustained release hydrophilic matrices by melt granulation in a high-shear mixer.
J Pharm Pharm Sci. 2005;8(2):132-40.
PURPOSE: The objective of this work was to prepare theophylline sustained release matrix tablets based on the combination of hydroxypropyl methylcellulose (HPMC K4M and K100M) and different meltable binders by melt granulation in a high-shear mixer. METHODS: Dissolution profiles of each formulation were compared to those of TheoDur 200 mg tablets and the mean dissolution time (MDT) and similarity factor (f2 factor) were calculated. The matrices swelling behavior was investigated by texture analysis. RESULTS: The results obtained show that the type of excipient influenced the drug release rate. In particular, the dissolution rate was delayed when lipophilic binders were used and only formulations containing Gelucire 50/13 or PEG 6000 with HPMC K4M had a profile similar to the commercial formulation. The release mechanism of theophylline from the formulations was described by Peppas's equation showing a non-Fickian release mechanism. The investigation of matrices swelling behavior showed that the gel layer thickness increased continuously over the time period studied. Moreover, a correlation between gel layer thickness and strength with the percentage released was found. CONCLUSIONS: These results suggest that melt granulation could be an easy and fast method to formulate sustained release tablets. [Abstract/Link to Full Text]

Brocks DR, Spencer TJ, Shayeganpour A
A sensitive and specific high performance liquid chromatographic assay for milrinone in rat and human plasma using a commercially available internal standard and low sample volume.
J Pharm Pharm Sci. 2005;8(2):124-31.
PURPOSE: To develop an alternate high performance liquid chromatographic method (HPLC) for the analysis of the positive inotropic agent, milrinone, in rat and human plasma. METHODS: To plasma samples (0.1 mL), containing milrinone and the commercially available internal standard, amrinone, were added 0.15 mL of water and 0.35 mL of acetonitrile. Tubes were briefly vortex-mixed and centrifuged. The supernatant was transferred to clean tubes and 3 mL of methanol: diethyl ether (5:95) was added. The tubes were vortex mixed, centrifuged, and reconstituted with the mobile phase and injected into the HPLC. Separation was accomplished using a reverse phase chromatography using C18 analytical column, and detection was afforded by monitoring the eluent at an ultraviolet wavelength of 326 nm. RESULTS: Standard curves were highly linear over the range 10 to 10000 ng/mL (r2 >0.99). Recovery ranged from 52-69% over a 40-fold range of plasma concentrations from 50 to 2000 ng/mL. Intra- and inter-day coefficient of variation and mean error in were less than 20% at plasma concentrations ranging from 10 to 1000 ng/mL. The utility of the assay was demonstrated in a pharmacokinetic evaluation of milrinone in two rats given intravenous bolus doses. CONCLUSION: The developed assay was sensitive, specific and appropriate for monitoring milrinone in rat or human plasma samples. [Abstract/Link to Full Text]

Pari L, Amali DR
Protective role of tetrahydrocurcumin (THC) an active principle of turmeric on chloroquine induced hepatotoxicity in rats.
J Pharm Pharm Sci. 2005;8(1):115-23.
PURPOSE: Tetrahydrocurcumin (THC) is an antioxidative substance, which is derived from curcumin, the component of turmeric. In the present investigation, the effect of THC and curcumin against chloroquine (CQ) induced hepatotoxicity were studied in female Wistar rats. METHODS: On single oral administration of CQ (970 mg/kg body weight) the activities of serum marker enzymes namely aspartate transaminase, alanine transaminase and alkaline phosphatase and the levels of bilirubin were significantly increased with significant alterations of lipids in serum and lipidperoxidation markers such as thiobarbituric acid reactive substances (TBARS) and hydroperoxides in plasma and liver were also elevated in CQ treated rats. The levels of non-enzymic antioxidants (vitamin C, vitamin E and reduced glutathione) and enzymic antioxidants (superoxide dismutase, catalase and glutathione peroxidase) were also decreased in CQ treated rats. Administration of THC (80 mg/kg body weight) and curcumin (80 mg/kg body weight) for 8 days before and 7 days after single administration of CQ significantly decreased the activities of serum markers and lipids in serum. In addition, the level of TBARS and hydroperoxides were significantly decreased with significant increase in non-enzymic and enzymic antioxidants on treatment with THC and curcumin. The biochemical observation was supplemented by histopathological examination of liver section. The results of the study reveal that THC shows more pronounced protective effect than curcumin against CQ induced toxicity. [Abstract/Link to Full Text]

Prakash H, Ali A, Bala M, Goel HC
Anti-inflammatory effects of Podophyllum hexandrum (RP-1) against lipopolysaccharides induced inflammation in mice.
J Pharm Pharm Sci. 2005;8(1):107-14.
PURPOSE: Down-regulation of lipopolysaccharide (LPS) induced hyper-inflammatory response by non-toxic pharmacological agents acquires paramount importance for countering bacterial sepsis. Anti-inflammatory potential of aqueous extract of Podophyllum hexandrum, a plant well documented in Ayurvedic literature for various therapeutic purposes, was investigated. METHODS: In vivo studies were performed on Balb/c mice pre-treated with supra-lethal dose of LPS endotoxin (E.coli 055:B5) with or without treatment with P. hexandrum extract (RP-1). Mouse peritoneal macrophage cultures were used to understand ex vivo effects of RP-1 on LPS generated nitric oxide (NO), secretion of IFN-gamma, IL-6 and TNF-alpha. Griess assay and sandwich ELISA method were used to quantify inducible NO and cytokines respectively. RESULTS: Minimal dose of LPS that rendered 100% mortality to mice was found to be 450 microg/kg b.w. Administration of RP-1 (200 mg/kg b.w., i.p.) one hour before lethal LPS treatment (0.5 mg/kg b.w.) rendered maximum (78%) survival. Ex vivo study revealed that RP-1 (50 microg/ml) treatment to peritoneal macrophages inhibited LPS (5 microg/ml) induced nitrite generation to 37%, IFN-gamma secretion to 5%, IL-6 secretion to 50% and TNF-alpha secretion to 50 % of LPS treated control values. CONCLUSION: This study has demonstrated anti-inflammatory potential of aqueous extract of P. hexandrum. [Abstract/Link to Full Text]

Amanlou M, Dadkhah F, Salehnia A, Farsam H, Dehpour AR
An anti-inflammatory and anti-nociceptive effects of hydroalcoholic extract of Satureja khuzistanica Jamzad extract.
J Pharm Pharm Sci. 2005;8(1):102-6.
PURPOSE: Satureja khuzistanica Jamzad (Lamiaceae) is an endemic plant that widely distributed in the southern parts of Iran. This plant has been used as analgesic and antiseptic among the inhabitants of southern parts of Iran. METHODS: The Satureja khuzistanica hydroalcoholic extract was prepared and its anti-inflammatory and anti-nociceptive effects were investigated using the carrageenan-induced rat paw edema and formalin test. RESULTS: A similar anti-inflammatory activity was seen between S. khuzistanica hydroalcoholic extract (150 mg/kg; i.p.) and indomethacin (4 mg/kg; i.p.) in carrageenan test. The extract showed anti-nociceptive activity in a dose-dependent (10-150 mg/kg; i.p.) manner at the second phase of formalin test which was comparable with morphine (3 mg/kg; i.p.). CONCLUSION: This study confirms that anti-inflammatory and anti-nociceptive properties of S. khuzistanica are comparable to those of indomethacin and morphine. Presence of flavonoids, steroids, essential oil, mainly carvacrol and tannin might be responsible for anti-inflammatory and anti-nociceptive activities of this plant. [Abstract/Link to Full Text]

Moon PD, Na JJ, Jeong HJ, Hong SH, Kim SJ, Chae HJ, Kim HR, Choi JO, Lee SH, Shin JY, Kim HM
Inhibitory effect of Gamibojungikgitang extract on mast cell-mediated allergic reaction in murine model.
J Pharm Pharm Sci. 2005;8(1):94-101.
PURPOSE: Gamibojungikgitang (GBIT) is an Oriental herbal prescription medication, which has been commonly used to treat allergic rhinitis in far Eastern countries including Korea, China, and Japan. Additionally, GBIT effectively treats ovarian cysts and improves ovarian functions by regulating both endocrine and metabolic activities. However, it has not been cleared how it prevents allergic diseases in experimental model. Here we report the effect of GBIT on mast cell-mediated allergic reactions. METHODS: The anti-anaphylactic effect of GBIT extract was studied against compound 48/80-induced systemic anaphylactic shock model in mice. Rat Peritoneal Mast Cells (RPMCs) were used to investigate the effect of GBIT extract on histamine release induced by compound 48/80. Passive cutaneous anaphylaxis activated by anti-dinitrophenyl IgE was used to know the effect of GBIT extract. In addition, human mast cell line HMC-1 cells culture supernatants that GBIT extract pretreated were assayed for IL-6 protein levels by enzyme-linked immunosorbent assay method. RESULTS: GBIT extract dose dependently inhibited compound 48/80-induced systemic anaphylactic shock. When GBIT extract was given as pretreatment at concentrations ranging 0.01-1 mg/ml, the histamine release from rat peritoneal mast cells induced by compound 48/80 was reduced in a dose-dependent manner. GBIT extract also inhibited passive cutaneous anaphylaxis activated by anti-dinitrophenyl IgE. In addition, GBIT extract inhibited phorbol 12-myristate 13-acetate + A23187-induced interleukin-6 secretion from human mast cell line HMC-1 cells. CONCLUSION: These results suggest a potential role for GBIT extract as a source of anti-anaphylactic agents for allergic disorders. [Abstract/Link to Full Text]

Gohel MC, Jogani PD
A review of co-processed directly compressible excipients.
J Pharm Pharm Sci. 2005;8(1):76-93.
Direct compression is the preferred method for the preparation of tablets. The present review outlines the importance of the functionality of the directly compressible adjuvants in the formulation of tablets. The co-processing is the most widely explored method for the preparation of directly compressible adjuvants because it is cost effective and can be prepared in-house based on the functionality required. Hence, the present review focuses on the properties of the co-processed directly compressible adjuvants available in the market. [Abstract/Link to Full Text]

Amaral Pde A, Bergold AM, Eifler-Lima VL, Santos EM, Oliveira ER, Campos-Buzzi F, Cechinel-Filho V
Antinociceptive effects of some synthetic delta-valerolactones.
J Pharm Pharm Sci. 2005;8(1):69-75.
PURPOSE: A series of delta-valerolactones has been synthesized in good yields, by reaction of ethyl acetoacetate with several aldehydes in presence of LDA. METHODS AND RESULTS: The in vivo analgesic activity of these compounds has been evaluated. The results indicate that the lactones synthesized showed an important antinociceptive effect at 10 mg/kg, administered intraperitoneally in mice, which significantly inhibited the abdominal constrictions induced by acetic acid when compared to acetylsalicylic acid and acetaminophen in the same dose, and increased significantly the thermal sensibility at the hot-plate method, although they were less effective than morphine in the same assay. CONCLUSIONS: The antinociceptive models employed here reveal a potential analgesic effect of the delta-valerolactones synthesized. Further investigations are needed to discern which mechanism of action is concern. [Abstract/Link to Full Text]

Emendörfer F, Emendörfer F, Bellato F, Noldin VF, Niero R, Cechinel-Filho V, Cardozo AM
Evaluation of the relaxant action of some Brazilian medicinal plants in isolated guinea-pig ileum and rat duodenum.
J Pharm Pharm Sci. 2005;8(1):63-8.
PURPOSE: The present study aimed to evaluate the possible antispasmodic activity in vitro of methanolic extracts (ME) of six Brazilian medicinal plants. METHODS: The extracts were evaluated on isolated guinea-pig ileum and rat duodenum preparations. RESULTS: Rubus imperialis, Maytenus robusta, Ipomoea pes caprae and Epidendrum mosenii did not inhibit the contractile response elicited by acetylcholine on guinea-pig ileum. On the other hand, ME from Calophyllum brasiliense and Cynara scolymus exhibited significant inhibitory activity for the contractile response elicited by acetylcholine on guinea-pig ileum and on rat duodenum in a noncompetitive and concentration-dependent manner. CONCLUSIONS: The current study suggests that, of the six medicinal plants evaluated, only the ME of Calophyllum brasiliense and Cynara scolymus show probable antispasmodic activity, confirming and justifying their use in folk medicine for the treatment of intestinal disorders. [Abstract/Link to Full Text]

Mashru R, Sutariya V, Sankalia M, Sankalia J
Transbuccal delivery of lamotrigine across porcine buccal mucosa: in vitro determination of routes of buccal transport.
J Pharm Pharm Sci. 2005;8(1):54-62.
PURPOSE: To determine the major routes of buccal transport of lamotrigine and to examine the effects of pH on drug permeation. METHODS: Transbuccal permeation of lamotrigine across porcine buccal mucosa was studied by using in-line Franz type diffusion cell at 37 degrees C. The permeability of lamotrigine was determined at pH 4.0 to 9.0. The permeability of unionized (Pu) and ionized (Pi) species of drug were calculated by fitting the data to a mathematical model. Lamotrigine was quantified by using the HPLC method. RESULTS: The steady state flux increased linearly with increasing the donor concentration (r2 = 0.9639) at pH 7.4. The permeability coefficient and the partition coefficient of the drug increased with increasing the pH. The values of Pu and Pi were 0.7291 x 10(-5) cm/sec and 0.2500 x 10(-5) cm/sec, respectively. The observed permeability coefficients and the permeability coefficients calculated from mathematical model at various pH showed good linearity (r(2) = 0.9267). The total permeability coefficient increased with increasing the fraction of unionized form of the drug. CONCLUSION. Lamotrigine permeated through buccal mucosa by a passive diffusion process. The partition coefficient and pH dependency of drug permeability indicated that lamotrigine was transported mainly via the transcellular route by a partition mechanism. [Abstract/Link to Full Text]

Tassaneeyakul W, Kittiwattanagul K, Vannaprasaht S, Kampan J, Tawalee A, Puapairoj P, Tiamkao S, Juthagridsada S, Kukongviriyapan V, Tassaneeyakul W
Steady-state bioequivalence study of clozapine tablet in schizophrenic patients.
J Pharm Pharm Sci. 2005;8(1):47-53.
PURPOSE: To compare the bioavailability of two clozapine formulations (100 mg Clozaril tablet from Novartis Pharmaceuticals UK Ltd., UK, as a Reference formulation and 100 mg Cloril tablet from Atlantic Laboratories Corp., Ltd., Thailand, as a Test formulation). The present study was conducted under real-life conditions in schizophrenic patients using a steady-state, multiple-dose, randomized crossover design to avoid the risk of adverse effects in healthy volunteers and pharmacokinetic difference between single and multiple-dose of the drug. METHODS: The subjects received 100 mg bid of either the Reference formulation or the Test formulation for 7 days. At day-7 of each study phase, blood samples were collected at 0, 0.5, 1, 1.5, 2, 2.5, 3, 4, 6, 8, 10 and 12 h after drug administration. Plasma was separated and stored at -80 degrees C until assay. The plasma concentration of clozapine was determined by high performance liquid chromatography. Pharmacokinetic parameters were calculated from the observed plasma-concentration time profiles. The bioequivalence between the two formulations was assessed by calculating individual peak plasma concentrations (Cmax) and area under the concentration-time curve (AUC(0-12 h)) ratios. RESULTS: All subjects well tolerated both clozapine formulations. No serious side effects were reported. The Tmax, terminal half-life and the total plasma clearance of clozapine (uncorrected for bioavailability) observed in the present study were comparable to those observed in other previous reports. All of the pharmacokinetic parameters investigated in the present study calculated from the subjects after administration of Test and Reference formulations were close. The 90% confident interval for the ratio of means for the lnCmax (0.9784-1.0622) and lnAUC(0-12h) (0.9559-1.0441) are within the guideline range of bioequivalence (0.80 to 1.25). CONCLUSION: The result demonstrated that the Test formulation was bioequivalent to the Reference formulation (Clozaril) when orally administered in schizophrenic patients, in terms of both the rate and extent of absorption. [Abstract/Link to Full Text]

Venkatesan N, Thiyagarajan V, Narayanan S, Arul A, Raja S, Vijaya Kumar SG, Rajarajan T, Perianayagam JB
Anti-diarrhoeal potential of Asparagus racemosus wild root extracts in laboratory animals.
J Pharm Pharm Sci. 2005;8(1):39-46.
PURPOSE: Asparagus racemosus Wild root has been used traditionally in Ayurveda for the treatment of diarrhoea and dysentery. However, the claims of Ayurveda need to be validated by a suitable experimental model. Therefore, the present study was undertaken to evaluate the effect of ethanol and aqueous extracts of Asparagus racemosus for its antidiarrhoeal potential against several experimental models of diarrhoea in Albino Wistar rats. METHODS: The antidiarrhoeal activity of ethanol and aqueous extracts of Asparagus racemosus root was evaluated using castor oil-induced diarrhoea model in rats. Further, we evaluated the effect of ethanol and aqueous extracts on gastrointestinal tract motility after charcoal meal administration and PGE2 induced intestinal fluid accumulation (enteropooling). Loperamide was used as positive control. RESULTS: The plant extracts showed significant (P < 0.05) inhibitor activity against castor oil induced diarrhoea and PGE2 induced enteropooling in rats when tested at 200 mg/kg. Both extracts also showed significant (P < 0.001) reduction in gastrointestinal motility in charcoal meal test in rats. CONCLUSION: The results point out the possible anti-diarrhoeal effect of the plant extracts and substantiate the use of this herbal remedy as a non-specific treatment for diarrhoea in folk medicine. [Abstract/Link to Full Text]

Mutalik S, Udupa N
Formulation development, in vitro and in vivo evaluation of membrane controlled transdermal systems of glibenclamide.
J Pharm Pharm Sci. 2005;8(1):26-38.
PURPOSE: The objective of the present study was to develop the membrane controlled transdermal systems of glibenclamide and to evaluate with respect o various in vitro and in vivo parameters. METHODS: The membrane moderated transdermal systems were prepared using drug containing carbopol gel as reservoir and ethyl cellulose, Eudragit RS-100, Eudragit RL-100 and Ethylene vinyl acetate (EVA) (2%, 9% and 19% vinyl acetate content) rate controlling membranes. The possible interaction between drug and polymer was studied by IR spectroscopy, DSC and HPTLC analysis. The formulations were subjected to various physicochemical studies, in vitro drug release studies and permeation studies through mouse skin. The blood glucose reducing hypoglycemic activity of the systems was studied in both normal and diabetic mice. Various biochemical parameters and histopathological studies were carried out after treating the diabetic mice for 6 weeks. Skin irritation tests, oral glucose tolerance test and pharmacokinetic evaluations were carried out in mice. RESULTS: The results suggested no interaction between drug and polymer. Variations in drug release/permeation profiles among the formulations containing different rate controlling membranes were observed. The scanning electron microscopic studies of EVA membranes demonstrated no changes in the surface morphology after in vitro skin permeation studies. The system with EVA rate controlling membrane (with 19% vinyl acetate) was selected for in vivo experiments. The transdermal system produced better improvement with respect to hypoglycemic activity, glucose tolerance test, all the tested biochemical, histopathological and pharmacokinetic parameters compared to oral administration, and exhibited negligible skin irritation. CONCLUSION: The present study shows that membrane controlled transdermal systems of glibenclamide exhibited better control of hyperglycemia and more effectively reversed the diabetes mellitus complications than oral glibenclamide administration in mice. [Abstract/Link to Full Text]

Nokhodchi A, Javadzadeh Y, Siahi-Shadbad MR, Barzegar-Jalali M
The effect of type and concentration of vehicles on the dissolution rate of a poorly soluble drug (indomethacin) from liquisolid compacts.
J Pharm Pharm Sci. 2005;8(1):18-25.
PURPOSE: For poorly soluble, highly permeable (Class II) drugs, such as indomethacin, the rate of oral absorption is often controlled by the dissolution rate in the gastrointestinal tract. Therefore together with the permeability, the solubility and dissolution behaviour of a drug are key determinants of its oral bioavailability. The object of the present study is to increase dissolution rate of indomethacin using liquisolid compacts. METHODS: Several formulations of liquisolid compacts containing various ratios of drug: propylene glycol (ranging from 1:1 to 1:4) was prepared. In this study the ratio of microcrystalline cellulose (carrier) to silica (coating powder material) was 20 in all formulations. The dissolution behaviour of indomethacin from liquisolid compacts and conventional formulations was investigated at different pHs (1.2 and 7.2). RESULTS: The results showed that liquisolid compacts demonstrated considerably higher drug dissolution rates than those of conventionally made capsules and directly compressed tablets containing indomethacin. This was due to increased wetting properties and surface of drug available for dissolution. Also it has been shown that the fraction of molecularly dispersed drug (FM) in the liquid medication of liquisolid systems was directly proportional to their indomethacin dissolution rates (DR). An attempt was made to correlate the percentage drug dissolved in 10-min with the solubility of indomethacin in different vehicles. A plot of the percentage drug dissolved against the solubility of indomethacin showed that the amount of drug dissolved increased linearly (correlation coefficient of 0.9994 and 0.996 at pH 7.2 and 1.2 respectively) with an increase in solubility of indomethacin in the vehicles. CONCLUSION: The liquisolid compacts technique can be a promising alternative for the formulation of water insoluble drugs, such as indomethacin into rapid release tablets. [Abstract/Link to Full Text]

Rana V, Babita K, Goyal D, Tiwary A
Sodium citrate cross-linked chitosan films: optimization as substitute for human/rat/rabbit epidermal sheets.
J Pharm Pharm Sci. 2004 Dec 20;8(1):10-7.
PURPOSE. To prepare chitosan films that shall be capable of simulating in vitro permeation of polar (5-FU) and non polar (indomethacin) drugs across rat/rabbit/human epidermis. METHOD: Statistical designs were utilized to identify the formulation and process variables that significantly influenced permeation of both drugs across chitosan films cross-linked with sodium citrate (Nacit). In addition, atomic absorption spectroscopy for Na+ and differential scanning calorimetry of these films were performed for understanding the cross-linking behaviour. RESULTS: Concentration of chitosan, cross-linking time and concentration of cross-linking agent significantly influenced the in vitro flux of both drugs. Na+ content in films cross-linked with Nacit solutions without adjustment of pH was found to increase whereas, DeltaH of cross-linking endothermic transition decreased. However, after dipping these films in phosphate buffer (pH 7.4) both Na+ and DeltaH was found to decrease. Such a decrease was not observed when films cross-linked with sodium citrate solutions after adjustment to pH 5 were dipped in phosphate buffer (pH 7.4). The in vitro permeation of both drugs across latter films was significantly less. CONCLUSION: Adjustment of Nacit solution pH to 5 produced ionic complexes that were resistant to alkaline pH. Chitosan films can be cross-linked with Nacit to simulate in vitro permeation of polar and non polar drugs across animal/human epidermis. [Abstract/Link to Full Text]

Ng SP, Wong KY, Zhang L, Zuo Z, Lin G
Evaluation of the first-pass glucuronidation of selected flavones in gut by Caco-2 monolayer model.
J Pharm Pharm Sci. 2004 Dec 20;8(1):1-9.
PURPOSE. Four flavones, namely Apigenin, Baicalein, Chrysin and Luteolin, were selected for study and comparison of their absorption and metabolism in gut using the in vitro Caco-2 monolayer model. METHODS. Transport of the four flavones in the Caco-2 monolayer model was studied in both Apical-to-Basolateral and Basolateral-to-Apical directions. RESULTS. All of the selected flavones were able to pass through the Caco-2 cell monolayer with no significant efflux. The permeability coefficients of the four compounds were all greater than 10(-6) cm/sec and those of Apigenin and Baicalein were even greater than 10(-5) cm/sec. Glucuronides of the tested flavones were all formed in the Caco-2 cell monolayer model and a structure-activity relationship has been proposed for this glucuronidation. In addition, Apical-to-Basolateral transport studies were performed in Caco-2 models pre-treated with Chrysin, an UGT inducer. Quantities of the corresponding glucuronides formed were all significantly higher in Chrysin-treated groups than the controls. CONCLUSIONS. It demonstrated that all selected flavones were substrates of the UGT isoforms that are inducible by Chrysin. [Abstract/Link to Full Text]

McQuarrie S, Mercer J, Syme A, Suresh M, Miller G
Preliminary results of nanopharmaceuticals used in the radioimmunotherapy of ovarian cancer.
J Pharm Pharm Sci. 2005;7(4):29-34.
PURPOSE: The treatment of late stage ovarian cancer presents an unmet clinical need for women around the world. A multistep radioimmunotherapeutic (RIT) approach, exploiting the combination of a bispecific monoclonal antibody (BsMAb) with 90Y labelled biotinylated long-circulating liposomes was tested as a potential adjuvant treatment for epithelial ovarian carcinomatosis in an attempt to meet this need. This approach was used to overcome some of the major obstacles associated with conventional strategies, in particular, to increase the amount of radioactivity delivered to the tumor site compared with conventional monoclonal antibody (MAb) radionuclide delivery. We hypothesize that sequential intraperitoneal administration of the targeting and therapeutic moieties provides the basis for an enhanced therapeutic ratio. METHODS: A BsMAb, with anti-CA 125 and anti-biotin epitopes was engineered for use with PEGylated liposomes coated with biotin to deliver the cytotoxic radionuclide 90Y to tumor sites. An in vivo therapy trial was used to test this RIT protocol with Balb/c nude mice (n=29) xenografted with the NIH:OVCAR-3 (CA 125+) human ovarian cancer cell line. RESULTS: A median tumor growth delay of 91 days for the combined treatment group versus 77.7 days for the control group was observed. CONCLUSION: An ongoing tumor growth delay/control study using this model has indicated an appreciable delay in progress of tumor and ascites development in treated vs. control populations. [Abstract/Link to Full Text]

Zwiorek K, Kloeckner J, Wagner E, Coester C
Gelatin nanoparticles as a new and simple gene delivery system.
J Pharm Pharm Sci. 2005;7(4):22-8.
PURPOSE: The aim of this study was to evaluate cationized gelatin nanoparticles as biodegradable and low cell toxic alternative carrier to existing DNA delivery systems. METHODS: Native gelatin nanoparticles were produced using a two step desolvation method. In order to bind DNA by electrostatic interactions onto the surface of the particles, the quaternary amine cholamine was covalently coupled to the particles. The modified nanoparticles were loaded with different amounts of plasmid in varying buffers and compared to polyethyleneimine-DNA complexes (PEI polyplexes) as gold standard. Transfection ability of the loaded nanoparticles was tested on B16 F10 cells. Additionally, the cell toxicity of the formulations was monitored. RESULTS: Different setups resulted in efficient gene delivery displayed by exponential increase of gene expression. The gene expression itself occurred with a certain delay after transfection. In contrast to PEI polyplexes, cationized gelatin nanoparticles almost did not show any significant cytotoxic effects. CONCLUSIONS: Cationized gelatin nanoparticles have shown the potential of being a new effective carrier for nonviral gene delivery. The major benefit of gelatin nanoparticles is not only the very low cell toxicity, but also their simple production combined with low costs and multiple modification opportunities offered by the matrix molecule. [Abstract/Link to Full Text]

Zillies J, Coester C
Evaluating gelatin based nanoparticles as a carrier system for double stranded oligonucleotides.
J Pharm Pharm Sci. 2005;7(4):17-21.
PURPOSE: Surface modified gelatin nanoparticles were tested as a potential carrier system for double stranded DNA and RNA oligonucleotides. The results will be discussed with regard to former experiments conducted with single stranded oligonucleotides. METHODS: Gelatin nanoparticles were prepared by a two step desolvation method and surface modified by the covalent coupling of a quaternary amine to obtain a permanent positive net charge. Oligonucleotide loading was conducted in three different media applying 50 microg oligonucleotide per mg nanoparticles in total. Five batches of nanoparticles varying in size and zeta potential (zeta) were tested. The zeta potentials were determined under enforced ionic conditions in a 10 mmol sodium chloride solution at pH 7.0. The separation of unbound oligonucleotides and gelatin nanoparticles was achieved by centrifugation. Free oligonucleotide was determined UV-spectrophotometrically (260 nm) in the supernatant. RESULTS: It could be shown that up to 50 microg nucleic acid per mg nanoparticles can be bound depending on the particle's zeta potential and the chosen incubation medium. CONCLUSIONS: The results suggest that the proposed procedure allows a successful drug loading of double stranded oligonucleotides onto to the surface of accordingly modified gelatin nanoparticles. [Abstract/Link to Full Text]

Guttikonda S, Wang W, Suresh M
Molecular zipper assays: a simple homosandwich with the sensitivity of PCR.
J Pharm Pharm Sci. 2004;7(4):7-16.
PURPOSE: The purpose of this study is to develop a simple and inexpensive method for detection of viral load or antigens present in the body fluids as for diagnosis or monitoring of infectious diseases. For example, in case of viral infection, nucleic acid based quantitative PCR/RTPCR are sensitive in measuring viral load to follow the course of therapy or infection. The key limitations of such assays include the need for sample extraction, susceptibility to inhibitors, and high cost. METHODS: A molecular zipper assay based on the simple homosandwich concept for repeated epitopes was developed where the analyte or virus is sandwiched between the same antibodies for detection. A comparative study of the lower limit of detection of M13 model virus was performed with various substrates. RESULT: Homosandwich molecular zipper assay captured the model virus with high avidity resisting multiple rounds of washing. Detection of the virus by enzyme labeled MAb in combination with chemiluminescent substrates provided practical assay sensitivities of 7-15 phages and a theoretical detection sensitivity of one virus particle. CONCLUSION: The significance of our results on the molecular zipper assay relates to the development of ultrasensitive pathogen assays at low cost. Such assays could be developed for pathogenic bacteria and viruses, especially HIV & HCV viruses, which are ravaging impoverished continents of Africa, Asia and Latin America. [Abstract/Link to Full Text]

Adams M, Kwon GS
Spectroscopic investigation of the aggregation state of amphotericin B during loading, freeze-drying, and reconstitution of polymeric micelles.
J Pharm Pharm Sci. 2004;7(4):1-6.
PURPOSE: To investigate the relative aggregation state of amphotericin B (AmB) during loading and reconstitution of polymeric micelles. METHODS: Hexanoate and stearate derivatives of PEO-b-p (L-Asp) were prepared. The polymers and AmB were dissolved in methanol (MeOH). Milli-Q water was then added slowly, and the MeOH was removed via rotary evaporation. The solutions were freeze-dried in the presence of trehalose. During micelle preparation, the aggregation state of AmB was assessed using absorption spectroscopy. Upon reconstitution, the samples were analyzed using vapor-pressure osmometry, size-exclusion chromatography (SEC), and absorption spectroscopy. The absorption spectrum of AmB in the presence of the block copolymers was compared to that of AmB alone under the same conditions. RESULTS: AmB was loaded into micelles prepared from acyl derivatives of PEO-b-p (L-Asp). Absorption spectroscopy indicated that the aggregation state was preserved during the loading process. AmB exists in a self-aggregated state in polymeric micelles containing hexanoate ester cores and in a relatively monomeric state in polymeric micelles containing stearate ester cores. Vapor-pressure osmometry confirmed the isotonicity of the formulations, while SEC indicated that the micelles were approximately 10(6) g/mol. CONCLUSIONS: Depending on the polymer structure and assembly conditions, it is possible to encapsulate AmB in a relatively nonaggregated or aggregated state in micelles prepared from acyl derivatives of PEO-b-p (L-Asp). In polymeric micelles containing stearate side chains, AmB was loaded in a nearly monomeric state, possibly due to interaction with the stearate side chains. The final aggregation state of the drug is preserved during lyophilization and reconstitution of polymeric micelles prepared by a novel solvent evaporation procedure. [Abstract/Link to Full Text]

Barillaro V, Bertholet P, Henry de Hassonville S, Ziemon E, Evrard B, Delattre L, Piel G
Effect of acidic ternary compounds on the formation of miconazole/cyclodextrin inclusion complexes by means of supercritical carbon dioxide.
J Pharm Pharm Sci. 2004 Nov 30;7(3):378-88.
PURPOSE: The aim of the study is to evaluate the effect of different acidic compounds on the inclusion of miconazole (MICO) in several cyclodextrins (CDs) using supercritical carbon dioxide (SCCO(2) ) processing. METHODS: Physical mixtures were processed by SCCO(2) at 30 MPa, 125 degrees C during 60 minutes in a static mode to produce inclusion complexes. The inclusion complexes were characterized by differential solubility, Fourier transform infrared spectroscopy (FT-IR) and dissolution test. RESULTS: The best inclusion yields were achieved with the combination of MICO base and HPgammaCD with or without acids. Maleic and fumaric acids influenced the MICO inclusion differently in function of their conformation. During the process, a miconazole salt was observed with maleic acid and characterized by thermal analysis and mass spectrometry. The kinetics inclusion followed a saturation-type shape curve. FT-IR confirmed the presence of genuine inclusion complexes. The complexes MICO base/HPgammaCD/(L-tartaric acid) enhanced the dissolution rates of MICO more than the corresponding physical mixtures did. Lastly, the stability study revealed that the complexes were stable. CONCLUSIONS: The formation of stable complexes between MICO and CDs is possible using SCCO(2). Moreover an acidic ternary compound is able to modify the formation of the complex. The inclusion complexes, which show better dissolution profiles than those with the corresponding physical mixtures, could lead to an increase of the oral bioavailability of MICO. [Abstract/Link to Full Text]

Recent Articles in Journal of Drug Targeting

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Recent Articles in Drug Metabolism and Pharmacokinetics

Sakaeda T
MDR1 genotype-related pharmacokinetics: fact or fiction?
Drug Metab Pharmacokinet. 2005 Dec;20(6):391-414.
Multidrug resistant transporter MDR1/P-glycoprotein, the gene product of MDR1, is a glycosylated membrane protein of 170 kDa, belonging to the ATP-binding cassette superfamily of membrane transporters. A number of various types of structurally unrelated drugs are substrates for MDR1, and MDR1 and other transporters are recognized as an important class of proteins for regulating pharmacokinetics. The first investigation of the effects of MDR1 genotypes on pharmacotherapy was reported in 2000; a silent single nucleotide polymorphism (SNP), C3435T in exon 26, was found to be associated with the duodenal expression of MDR1, and thereby the plasma concentration of digoxin after oral administration. In the last 5 years, clinical studies have been conducted around the world on the association of MDR1 genotype with MDR1 expression and function in tissues, and with the pharmacokinetics and pharmacodynamics of drugs; however, there are still discrepancies in the results on C3435T. In 1995, a novel concept to predict in vivo oral pharmacokinetic performance from data on in vivo permeability and in vitro solubility has been proposed, and this Biopharmaceutical Classification System strongly suggested that the effects of intestinal MDR1 on the intestinal absorption of substrates is minimal in the case of commercially available oral drugs, and therefore MDR1 genotypes are little associated with the pharmacokinetics after oral administration. This review summarizes the latest reports for the future individualization of pharmacotherapy based on MDR1 genotyping, and attempts to explain discrepancies. [Abstract/Link to Full Text]

Hiratsuka M, Kudo M, Koseki N, Ujiie S, Sugawara M, Suzuki R, Sasaki T, Konno Y, Mizugaki M
A novel single nucleotide polymorphism of the human methylenetetrahydrofolate reductase gene in Japanese individuals.
Drug Metab Pharmacokinet. 2005 Oct;20(5):387-90.
The genetic polymorphisms of methylenetetrahydrofolate reductase (MTHFR) have been associated with increased toxicity of methotrexate (MTX), a folic acid antagonist that is widely used to treat cancer and immunosuppressive disorders such as rheumatoid arthritis. In this study, we analyzed all the exons and exon/intron junctions of the MTHFR gene from 200 Japanese individuals. We detected a novel single nucleotide polymorphism (SNP) 148C>T (Arg46Trp) in exon 1. The allele frequency of this polymorphism in the Japanese population appears to be extremely low (0.25%). [Abstract/Link to Full Text]

Kimura N, Masuda S, Tanihara Y, Ueo H, Okuda M, Katsura T, Inui K
Metformin is a superior substrate for renal organic cation transporter OCT2 rather than hepatic OCT1.
Drug Metab Pharmacokinet. 2005 Oct;20(5):379-86.
Although metformin, a cationic agent for type II diabetes, shows its pharmacological effect in the liver, the drug is mainly eliminated into urine. The tissue selectivity based on the function of drug transporters is unclear. In the present study, the transport of metformin was examined using HEK293 cells transiently transfected with five human renal organic ion transporter cDNAs. Human OCT1 and OCT2, but not OAT1, OAT3 or OCT2-A, stimulated the uptake. A kinetic analysis of metformin transport demonstrated that the amount of plasmid cDNA for transfection was also important parameter to the quantitative elucidation of functional characteristics of transporters, and both human and rat OCT2 had about a 10- and 100-fold greater capacity to transport metformin than did OCT1, respectively. In male rats, the mRNA expression level of rOCT2 in the whole kidneys was 8-fold greater than that of rOCT1 in the whole liver. The in vivo distribution of metformin in rats revealed that the expression level of renal OCT2 was a key factor in the control of the concentrative accumulation of metformin in the kidney. These findings suggest that metformin is a superior substrate for renal OCT2 rather than hepatic OCT1, and renal OCT2 plays a dominant role for metformin pharmacokinetics. [Abstract/Link to Full Text]

Matsui-Sakata A, Ohtani H, Sawada Y
Receptor occupancy-based analysis of the contributions of various receptors to antipsychotics-induced weight gain and diabetes mellitus.
Drug Metab Pharmacokinet. 2005 Oct;20(5):368-78.
OBJECTIVE: Among various adverse reactions of atypical antipsychotics, weight gain and impaired glucose tolerance are clinically significant. The aim of this study is to analyze quantitatively the contributions of various receptors to these antipsychotics-induced adverse reactions based on the receptor occupancy theory. METHODS: Two indices of antipsychotics-induced weight gain (the values estimated by a meta-analysis and the observed values in clinical trials) and the morbidity rate of type 2 diabetes mellitus during treatment with antipsychotics were taken from the literature. We calculated the estimated mean receptor occupancies of alpha1 adrenergic, alpha2 adrenergic, dopamine D2, histamine H1, muscarinic acetylcholine (mACh), serotonin 5-HT1A, 5-HT2A and 5-HT2C receptors by antipsychotics by using the pharmacokinetic parameters and receptor dissociation constants, and analyzed the correlation between the occupancies and the extent of adverse reactions as assessed using the aforementioned indices. RESULTS: There were statistically significant correlations between the estimated occupancies of H1 and mACh receptors and antipsychotics-induced weight gain estimated by meta-analysis (r(s) = 0.81 and r(s) = 0.83, respectively, p < 0.01). There were also statistically significant correlations between these receptor occupancies and observed weight gain in clinical trials (r(s) = 0.66 in each case, p < 0.01). The morbidity rate of type 2 diabetes mellitus was highly correlated with H1, mACh, and 5-HT2C receptor occupancies (r(s) = 0.90 in each case, p < 0.05). However, H1 receptor occupancy was also highly correlated with mACh receptor occupancy among antipsychotics, so that only one of them may be critically associated with the adverse reactions. Considering that these adverse reactions have not been reported for drugs with mACh receptor antagonistic action, other than antipsychotics, the H1 receptor may contribute predominantly to the antipsychotics-induced weight gain and diabetes mellitus. DISCUSSION/CONCLUSION: Model analysis based on receptor occupancy indicates that H1 receptor blockade is the primary cause of antipsychotics-induced weight gain and diabetes mellitus. [Abstract/Link to Full Text]

Funahashi T, Tanaka Y, Yamaori S, Kimura T, Matsunaga T, Ohmori S, Kageyama T, Yamamoto I, Watanabe K
Stimulatory effects of testosterone and progesterone on the NADH- and NADPH-dependent oxidation of 7beta-hydroxy-delta8-tetrahydrocannabinol to 7-oxo-delta8-tetrahydrocannabinol in monkey liver microsomes.
Drug Metab Pharmacokinet. 2005 Oct;20(5):358-67.
Microsomal alcohol oxygenase catalyzes the stereoselective oxidation of 7alpha- and 7beta-hydroxy-delta8-tetrahydrocannabinol (7alpha- and 7beta-hydroxy-delta8-THC) to 7-oxo-delta8-THC in monkey liver, and the activity for 7beta-hydroxy-delta8-THC is relatively higher than that for 7alpha-hydroxy-delta8-THC. We previously reported that purified P450JM-E, assumed to be CYP3A8, is a major enzyme responsible for the oxidation of 7-hydroxy-delta8-THC to 7-oxo-delta8-THC in monkey liver and is capable of catalyzing the oxidative reaction by NADH as well as NADPH. In the present study, we demonstrated that some steroids such as testosterone and progesterone stimulated both the NADH- and NADPH-dependent conversions of 7beta-hydroxy-delta8-THC to 7-oxo-delta8-THC in monkey liver microsomes. Kinetic analyses revealed that both the NADH- and NADPH-dependent 7-oxo-delta8-THC formation showed sigmoid kinetics. Testosterone caused a decrease in S50 and an increase in V(max) for the NADH-dependent activity, and resulted in a decrease in S50 without changing the V(max) for the NADPH-dependent activity. On the other hand, NADH-dependent testosterone 6beta-hydroxylation activity showed Michaelis-Menten kinetics and was also inhibited by 7beta-hydroxy-delta8-THC, resulting in a decrease in V(max) with no effect on the K(m). NADPH-dependent testosterone 6beta-hydrozylation activity was also inhibited by 7beta-hydroxy-delta8-THC, resulting in a decrease in both S50 and V(max). In order to explain the metabolic interaction between 7beta-hydroxy-delta8-THC and testosterone, we propose a kinetic model involving at least three binding sites, for the mechanism of activation by testosterone. [Abstract/Link to Full Text]

Emoto C, Murase S, Sawada Y, Iwasaki K
In vitro inhibitory effect of 1-aminobenzotriazole on drug oxidations in human liver microsomes: a comparison with SKF-525A.
Drug Metab Pharmacokinet. 2005 Oct;20(5):351-7.
1-Aminobenzotriazole (ABT) is extensively used as a non-specific cytochrome P450 (CYP) inhibitor. In this study, the inhibitory effect of ABT on CYP-dependent drug oxidations was investigated in human liver microsomes (HLM) and compared with that of SKF-525A, another non-specific inhibitor. The following probe activities for human CYP isoforms were determined using pooled HLM: phenacetin O-deethylation (CYP1A2); diclofenac 4'-hydroxylation (CYP2C9); S-mephenytoin 4'-hydroxylation, (CYP2C19); bufuralol 1'-hydroxylation (CYP2D6); chlorzoxazone 6-hydroxylation (CYP2E1); midazolam 1'-hydroxylation, nifedipine oxidation, and testosterone 6beta-hydroxylation (CYP3A). ABT had the strongest inhibitory effect on the CYP3A-dependent drug oxidations and the weakest effect on the diclofenac 4'-hydroxylation. SKF-525A potently inhibited the bufuralol 1'-hydroxylation, but weakly inhibited chlorzoxazone 6-hydroxylation. The inhibitory effects of ABT and SKF-525A were increased by preincubation in some probe reactions, and this preincubation effect was greater in ABT than in SKF-525A. The remarkable IC50 shift (> 10 times) by preincubation with ABT was observed on the phenacetin O-deethylation, chlorzoxazone 6-hydroxylation, and midazolam 1'-hydroxylation. In conclusion, ABT and SKF-525A had a wide range of IC50 values in inhibiting the drug oxidations by HLM with and without preincubation. [Abstract/Link to Full Text]

Fukuda T, Maune H, Ikenaga Y, Naohara M, Fukuda K, Azuma J
Novel structure of the CYP2D6 gene that confuses genotyping for the CYP2D6*5 allele.
Drug Metab Pharmacokinet. 2005 Oct;20(5):345-50.
We encountered DNA samples which showed a positive product using a long PCR-based method for the detection of CYP2D6*5, indicating deletion of the entire CYP2D6 gene, but the samples did not show a band related to CYP2D6*5 in either XbaI- or EcoRI-RFLP analysis. To achieve genotyping with accuracy, we performed a further genetic analysis to clarify the discrepancy. An unknown 1.6-kb insert was identified in a region downstream from the CYP2D6 stop codon where a specific primer was designed for long-PCR analysis for CYP2D6*5 genotyping. This finding suggested that the CYP2D6 gene might not be deleted in the samples even if a positive product was detected by the long-PCR method. Furthermore, the allelic frequency of this type was found to be approximately 0.3% (4 heterozygous/771 samples) in a Japanese population. In conclusion, we found a novel structure of the CYP2D6 gene, which might lead to incorrect genotyping for CYP2D6*5. Although the long PCR-based strategy for the detection of CYP2D6*5 has been widely used due to its usefulness and convenience, we recommend caution when adopting this method and propose re-evaluating the method for detecting CYP2D6*5. [Abstract/Link to Full Text]

Murakami M, Furuie H, Matsuguma K, Wanibuchi A, Kikawa S, Irie S
Pharmacokinetics and pharmacodynamics of landiolol hydrochloride, an ultra short-acting beta1-selective blocker, in a dose escalation regimen in healthy male volunteers.
Drug Metab Pharmacokinet. 2005 Oct;20(5):337-44.
OBJECTIVES: We conducted a randomized, double-blind, placebo-controlled study to evaluate the pharmacokinetics and pharmacodynamics of landiolol hydrochloride in a dose escalation regimen in healthy male volunteers. METHODS: We set two-dose escalation regimen (LM and MH groups) using three different doses [L (low): 0.03 mg/kg/min (1 min) loading-->0.01 mg/kg/min (10 min) continuous, M (medium): 0.06 mg/kg/min (1 min) loading-->0.02 mg/kg/min (10 min) continuous, H (high): 0.125 mg/kg/min (1 min) loading-->0.04 mg/kg/min (10 min) continuous]. Sixteen subjects were allocated randomly to the LM, MH, and placebo groups (n=6, 6, and 4, respectively). RESULTS: In both the LM and MH groups, the blood concentration of landiolol hydrochloride changed within a constant range from 2 minutes after initiation of administration to just before the higher dose escalation. By 2 minutes following the higher dose escalation, the concentration of landiolol hydrochloride reached C(max), and reached almost steady state levels until 6 minutes following administration of the higher dose. The t(1/2) of landiolol hydrochloride was 3.5 minutes. The heart rates and blood pressures of subjects administered landiolol hydrochloride decreased, but there were no adverse events in any subject. CONCLUSIONS: The concentration of landiolol hydrochloride rapidly reached steady state levels, and rapidly dissipated after completion of administration. [Abstract/Link to Full Text]

Harada S, Yamashita F, Hashida M
Estimation of in vivo percutaneous absorption of emedastine from bile excretion data using a deconvolution method.
Drug Metab Pharmacokinet. 2005 Oct;20(5):331-6.
In vivo percutaneous absorption of emedastine difumarate was investigated in rats and compared with rat skin in vitro. Since emedastine entering the systemic circulation is mostly excreted in bile, we first came up with the method of collecting bile with a minimal skin incision. In vivo skin permeation of the drug was estimated from biliary excretion data by deconvolution analysis. Prior to applying deconvolution analysis, it was confirmed that biliary excretion of emedastine was linear against its dose. When the in vivo permeation profile estimated by deconvolution was compared with the in vitro profile, the lag time for permeation was significantly shorter in vivo than in vitro, whereas the skin permeability coefficient was almost the same. If we presume a two-layer diffusion model, then this finding may primarily be due to the shorter diffusion length of the dermis. [Abstract/Link to Full Text]

Jin M, Shimada T, Shintani M, Yokogawa K, Nomura M, Miyamoto K
Long-term levothyroxine treatment decreases the oral bioavailability of cyclosporin A by inducing P-glycoprotein in small intestine.
Drug Metab Pharmacokinet. 2005 Oct;20(5):324-30.
We have noticed that the trough level of blood concentration of cyclosporin A (CyA) tends to be lower in patients receiving long-term oral levothyroxine (LTX) than in patients not receiving LTX. We confirmed this clinical observation in experiments using Wistar rats orally given LTX (8 microg/kg) or saline (control) for 3 weeks, followed by CyA (10 mg/kg). The LTX treatment had little effect on the blood concentrations of CyA after i.v. administration, whereas they were decreased significantly after p.o. administration. After p.o. administration, the value of the area under the blood concentration-time curve from 0 to 24 hr and the bioavailability of CyA in the LTX group were decreased to only about one-fifth and a quarter of those in the control group, respectively. After treatment with LTX, the expression levels of mdr1a, mdr1b and CYP3A2 mRNAs in the duodenum were markedly increased to about twice the control, but in jejunum, ileum and liver the expression levels were little changed. These findings suggest that the absorption of CyA, which occurs mainly from the upper intestine, is reduced as a result of efflux transport via P-glycoprotein induced by LTX. In conclusion, careful monitoring of CyA levels is required in the event of LTX administration to patients receiving immunotherapy with CyA. [Abstract/Link to Full Text]

Otagiri M
A molecular functional study on the interactions of drugs with plasma proteins.
Drug Metab Pharmacokinet. 2005 Oct;20(5):309-23.
The binding of drugs to plasma proteins, such as albumin and alpha1-acid glycoprotein (AGP) is a major determinant in the disposition of drugs. A topology analysis of drug binding sites on HSA and AGP was determined using various methods, including spectroscopy, QSAR, photoaffinity labeling and site directed mutagenesis. Recombinant albumin was found to be useful for rapidly identifying drug binding sites. The binding sites on AGP are not completely separated but are partially overlapped, and Trp, Tyr, Lys and His residues in the drug binding pockets play important roles in this process. Drug displacement is somewhat complex, due to the involvement of multiple effects. The reduced binding in uremic patients may be explained by a mechanism that involves a combination of direct displacement by free fatty acids as well as cascade effects of free fatty acids and unbound uremic toxins for significant inhibition in serum binding. Albumin-containing dialysate is useful for the extracorporeal removal of endogenous toxins and in the treatment of drug overdoses. Oxidized albumin is a useful biomarker for the quantitative and qualitative evaluation of oxidative stress. Interestingly, AGP undergoes a structural transition to a unique structure that differs from the native and denatured states, when it interacts with membranes. [Abstract/Link to Full Text]

Fukushima-Uesaka H, Saito Y, Maekawa K, Ozawa S, Hasegawa R, Kajio H, Kuzuya N, Yasuda K, Kawamoto M, Kamatani N, Suzuki K, Yanagawa T, Tohkin M, Sawada J
Genetic variations and haplotypes of CYP2C19 in a Japanese population.
Drug Metab Pharmacokinet. 2005 Aug;20(4):300-7.
Forty-eight single nucleotide variations, including 27 novel ones, were found in the 5'- regulatory region, all of the exons and their surrounding introns of CYP2C19 in 253 Japanese subjects (134 diabetic patients and 119 healthy volunteers). Identified novel variations were as follows: -2772G>A, 2767_-2760delGGTGAACA, -2720T>C, -2547delG, -2545G>T, -2545_-2544 delGC, and -2040C>T in the enhancer region; -778C>T, -777G>A, -529G>C, -189C>A, and -185A>G in the promoter region; 151A>G (S51G), 481G>C (A161P), 986G>A (R329H), 1078G>A (D360N), and 1119C>T (D373D) in the exons, and IVS1+128T>A, IVS3+163G>A, IVS4+271A>G, IVS5-49A>G, IVS6-210C>T, IVS6-196T>A, IVS6-32T>A, IVS7+84G>A, IVS7-174C>T, and IVS8+64C>T in the introns. Since we found no significant differences in the variation frequencies between healthy volunteers and diabetic patients, the data for all subjects were treated as one group in further analysis. The allele frequencies were 0.265 for IVS6-196T>A, 0.045 for -2772G>A and -2720T>C, 0.024 for -2040C>T, 0.014 for IVS7-174C>T, 0.010 for -529G>C, 0.006 for IVS1+128T>A and 481G>C (A161P), 0.004 for -2767_-2760delGGTGAACA and IVS6-210C>T, and 0.002 for the other 17 variations. In addition, the two known nonsynonymous single nucleotide polymorphisms, 681G>A (splicing defect, (*)2 allele) and 636G>A (W212X; (*)3 allele) were detected at 0.267 and 0.128 frequencies, respectively. No variation was detected in the known binding sites for constitutive androstane receptor and glucocorticoid receptor. Linkage disequilibrium analysis showed several close linkages of variations throughout the gene. By using the variations, thirty-one haplotypes of CYP2C19 and their frequencies were estimated. Our results would provide fundamental and useful information for genotyping CYP2C19 in the Japanese and probably other Asian populations. [Abstract/Link to Full Text]

Ebisawa A, Hiratsuka M, Sakuyama K, Konno Y, Sasaki T, Mizugaki M
Two novel single nucleotide polymorphisms (SNPs) of the CYP2D6 gene in Japanese individuals.
Drug Metab Pharmacokinet. 2005 Aug;20(4):294-9.
We analyzed all the exons and exon-intron junctions of the CYP2D6 gene from 286 Japanese individuals. We detected two novel single nucleotide polymorphisms (SNPs) 2556C>T in exon 5 (Thr261Ile) and 3835A>C in exon 8 (Lys404Gln). Both these SNPs showed a frequency of 0.002. [Abstract/Link to Full Text]

Murai T, Iwabuchi H, Ikeda T
Repeated glucuronidation at one hydroxyl group leads to structurally novel diglucuronides of steroid sex hormones.
Drug Metab Pharmacokinet. 2005 Aug;20(4):282-93.
Androgens (androsterone, dihydrotestosterone and testosterone) and estrogens (estradiol, estriol and estrone) were incubated with liver microsomes from rats, dogs, monkeys and humans in the presence of uridine diphosphoglucuronic acid (UDPGA), and the glucuronides produced were structurally characterized by liquid chromatography-tandem mass spectrometry. After 2-h incubation with dog liver microsomes, all substrates tested were converted (approximately 2-10%) to structurally novel diglucuronides, where two glucuronosyl groups are bound to a single hydroxyl group in tandem. Two-dimensional nuclear magnetic resonance spectroscopy unambiguously elucidated the chemical structures of the 3-O-diglucuronide of estrone and the 17-O-diglucuronide of testosterone isolated from the incubation mixture. Monkey and human liver microsomes were also found to have the activity to form this type of diglucuronide, albeit more slowly than the dog liver microsomes, but rat liver microsomes produced no detectable diglucuronides. The rate of formation of estrone 3-O-diglucuronide from the corresponding monoglucuronide in dog liver microsomes followed classical Michaelis-Menten kinetics at substrate concentrations from 50 to 1000 microM, with a K(m) value of 127.1 microM and a V(max) value of 47.0 pmol/min/mg protein. [Abstract/Link to Full Text]

Mudhakir D, Akita H, Khalil IA, Futaki S, Harashima H
Pharmacokinetic analysis of the tissue distribution of octaarginine modified liposomes in mice.
Drug Metab Pharmacokinet. 2005 Aug;20(4):275-81.
We recently found that octaarginine modified liposomes (R8-Lip) can be efficiently internalized by cultured cells. The purpose of the present study was to quantitatively determine the effect of R8-density on the tissue distribution of R8-Lip in mice, using their clearance as an index. R8 was introduced in the form of stearylated R8 (STR-R8). The liposomes were composed of cholesterol and egg phosphatidylcholine and were labeled with [(3)H]cholesteryl hexadecyl ether. Various densities of R8 (3%, 10% and 30%) containing liposomes were prepared with a diameter of approximately 70-80 nm. The tissue distribution of R8-Lip was determined after their i.v. administration into mice and the effect of R8-density on tissue distribution was compared with uptake clearance, the calculated tissue distribution divided by the area under the blood concentration-time course. As results, R8-Lip were more rapidly eliminated from circulating blood and distributed to many tissues, especially liver depending on the R8-density. However, the tissue uptake clearance represented similar value to that of positively charge liposomes. Based on these results, we conclude that the R8-dependent increase in R8-Lip in various tissues tested indicates that positive charge, but not PTD function derived from R8 predominantly responsible for the enhancement of tissue distribution. Therefore, it is suggested that topology control of R8 is important to exhibit the PTD function. [Abstract/Link to Full Text]

Aiba T, Susa M, Fukumori S, Hashimoto Y
The effects of culture conditions on CYP3A4 and MDR1 mRNA induction by 1alpha,25-dihydroxyvitamin D(3) in human intestinal cell lines, Caco-2 and LS180.
Drug Metab Pharmacokinet. 2005 Aug;20(4):268-74.
To evaluate an in vitro model suitable for investigating intestinal first-pass drug metabolism, CYP3A4 and MDR1 mRNA induction by 1alpha,25-dihydroxyvitamin D(3) (VD3) was examined in two human intestinal cell lines, Caco-2 and LS180, under various culture conditions. CYP3A4 mRNA expression was induced by 100 nM VD3 at levels between 234-549 times above normal in Caco-2 cells for 2 weeks and by 74-200 times above normal in LS180 cells for 2 days. The CYP3A4 induction effect of 250 nM VD3 was similar to or slightly higher than that of 100 nM VD3 in both Caco-2 and LS180 cells. Also, CYP3A4 was induced in Caco-2 and LS180 cells when they were cultured on a polystyrene plate slightly less than when they were cultured on a porous membrane. The increase in fetal bovine serum (FBS) content in the culture medium resulted in little or only slight increase of CYP3A4 induction in both Caco-2 and LS180 cells. MDR1 mRNA expression was marginally increased by VD3 in LS180 cells, but not in Caco-2 cells, and neither increased FBS content nor use of a porous membrane significantly affected MDR1 induction in LS180 cells. The transepithelial electrical resistance of LS180 cells was almost zero, whereas that of Caco-2 cells was high and was marginally decreased by VD3. These findings indicate that Caco-2 cells cultured on a porous membrane with 100 nM VD3 for 2 weeks may be used as a model to investigate the intestinal absorption and first-pass metabolism of drugs, while LS180 cells may be utilized to elucidate the mechanisms which regulate intestinal CYP3A4 mRNA expression. [Abstract/Link to Full Text]

Araya H, Tomita M, Hayashi M
The novel formulation design of self-emulsifying drug delivery systems (SEDDS) type O/W microemulsion II: stable gastrointestinal absorption of a poorly water soluble new compound, ER-1258 in bile-fistula rats.
Drug Metab Pharmacokinet. 2005 Aug;20(4):257-67.
The stabilization effect of the novel self-emulsifying drug delivery systems (SEDDS) type O/W microemulsion on the gastrointestinal absorption of a poorly water soluble new compound, ER-1258 was examined by bile-fistula model rats. In the components of this formulation, medium chain fatty acid triglyceride (MCT), diglyceryl monooleate (DGMO-C), polyoxyethylene hydrogenated castor oil 40 (HCO-40) and ethanol were used as an oil, a lipophilic surfactant, a hydrophilic surfactant and a solubilizer at the mixture ratio of 25/5/45/25 w/w%, respectively. The ratios of AUC in the non-treated rats to that in the bile-fistula rats were 5.1, 12.1 and 3.0 for the suspension, the oily solution and the SEDDS type O/W microemulsion, respectively. The risk from which the difference between individuals of the compound absorption amounts resulting from the flow of the bile secretion serves as the maximum was high in order of oily solution>suspension>SEDDS type O/W microemulsion. Therefore, it was verified that the SEDDS type O/W microemulsion was able to reduce this risk, compared with the other formulations. When short chain fatty acid triglyceride (Triacetin) was used as an oil, the similar effect was demonstrated in the formulation composed of sorbitan sesquioleate (SO-15) as a lipophilic surfactant and polyoxyethylene hydrogenated castor oil 60 (HCO-60) or polyoxyethylene 20 sorbitan monooleate (TO-10M) as a hydrophilic surfactant. [Abstract/Link to Full Text]

Araya H, Nagao S, Tomita M, Hayashi M
The novel formulation design of self-emulsifying drug delivery systems (SEDDS) type O/W microemulsion I: enhancing effects on oral bioavailability of poorly water soluble compounds in rats and beagle dogs.
Drug Metab Pharmacokinet. 2005 Aug;20(4):244-56.
We examined the design of the versatile novel self-emulsifying drug delivery systems (SEDDS) type O/W microemulsion formulation which enhances the oral bioavailability by raising the solubility of poorly water soluble compounds. Namely, seven kinds of poorly water soluble compounds such as disopyramide, ibuprofen, ketoprofen, tolbutamide, and other new compounds, as the model compounds were used to compare the plasma concentration profile of the compound following single oral administration of each compound to rats and beagle dogs as a solution, an oily solution, a suspension (or a powder), an O/W microemulsion, and a SEDDS type O/W microemulsion. And the enhancing effect of the SEDDS type O/W microemulsion on the gastrointestinal absorption of these compounds was evaluated. In the components of the SEDDS type O/W microemulsion, medium chain fatty acid triglyceride (MCT), diglyceryl monooleate (DGMO-C), polyoxyethylene hydrogenated castor oil 40 (HCO-40), and ethanol were used as an oil, a lipophilic surfactant, a hydrophilic surfactant, and a solubilizer, at the mixture ratio of 25/5/45/25 (w/w%), respectively. Thereby, to six kinds of the model compounds except disopyramide, the solubility was from 340 to 98,000 times that in water, and the AUCs in plasma concentration of the compound were equivalent to that of solution or O/W microemulsion administration, or was increased by 1.5 to 78 times that of suspension administration. Accordingly, this novel SEDDS type O/W microemulsion is the versatile, useful formulation which enhances the oral bioavailability by raising the solubility of poorly water soluble compounds. [Abstract/Link to Full Text]

Kato M, Chiba K, Horikawa M, Sugiyama Y
The quantitative prediction of in vivo enzyme-induction caused by drug exposure from in vitro information on human hepatocytes.
Drug Metab Pharmacokinet. 2005 Aug;20(4):236-43.
There have been no reports of the quantitative prediction of induction for drug-metabolizing enzymes in humans. We have tried to predict such enzyme induction in humans from in vitro data obtained using human hepatocytes. The in vitro and in vivo data on enzyme induction by inducers, such as rifampicin, phenobarbital and omeprazole, were collected from the published literature. The degree of enzyme induction in humans was compared with that predicted from in vitro data on human hepatocytes. Using the in vivo data, we calculated the hepatic intrinsic clearance of typical CYP substrates, such as midazolam and caffeine, before and after inducer treatment and estimated the induction ratios of hepatic intrinsic clearance following treatment. In the in vitro studies, the amount of mRNA or enzyme and enzyme activity in human hepatocytes, with or without an inducer, were compared and the induction ratios were estimated. The unbound mean concentration was taken as an index of drug exposure and the induction ratios in the in vivo and in vitro studies were compared. The unbound mean concentrations of inducers used in the in vitro studies were higher than those in the in vivo studies. The maximum induction ratios by inducers in the in vitro studies were higher than those in the in vivo studies. The induction ratio for rifampicin, omeprazole, troglitazone, dexamethasone and phenobarbital increased as the unbound mean concentration increased to reach a constant value. The induction of CYP3A and 1A was analyzed by the Emax model. The maximum induction ratio (Emax) and the concentration at half maximum induction (EC50) for rifampicin, omeprazole, troglitazone, dexamethasone and phenobarbital were 12.3, 0.847 micromol/L, 2.36, 0.225 micromol/L, 6.86, 0.002 micromol/L, 8.30, 9.32 micromol/L, and 7.62, 58.4 micromol/L, respectively. The Emax and EC50 of omeprazole for CYP1A were 12.02 and 0.075 micromol/L, respectively. The predicted induction ratio of all those inducers, except for omeprazole, based on the Emax and EC50 values obtained from the in vitro data were similar to the observed values. On the whole, a good correlation between the observed and predicted induction ratio of omeprazole was observed (r=0.768, p<0.05), although the predicted induction ratio was higher than the observed value. In conclusion, the present study suggests that it is possible to predict quantitatively the CYP3A enzyme induction from hepatocyte data. [Abstract/Link to Full Text]

Nakajima M, Yokoi T
Interindividual variability in nicotine metabolism: C-oxidation and glucuronidation.
Drug Metab Pharmacokinet. 2005 Aug;20(4):227-35.
Nicotine has roles in the addiction to smoking, replacement therapy for smoking cessation, as a potential medication for several diseases such as Parkinson's disease, Alzheimer's disease, and ulcerative colitis. The absorbed nicotine is rapidly and extensively metabolized and eliminated to urine. A major pathway of nicotine metabolism is C-oxidation to cotinine, which is catalyzed by CYP2A6 in human livers. Cotinine is subsequently metabolized to trans-3'-hydroxycotinine by CYP2A6. Nicotine and cotinine are glucuronidated to N-glucuronides mainly by UGT1A4 and partly by UGT1A9. Trans-3'-hydroxycotinine is glucuronidated to O-glucuronide mainly by UGT2B7 and partly by UGT1A9. Approximately 90% of the total nicotine uptake is eliminated as these metabolites and nicotine itself. The nicotine metabolism is an important determinant of the clearance of nicotine. Recently, advances in the understanding of the interindividual variability in nicotine metabolism have been made. There are substantial data suggesting that the large interindividual differences in cotinine formation are associated with genetic polymorphisms of the CYP2A6 gene. Interethnic differences have also been observed in the cotinine formation and the allele frequencies of the CYP2A6 alleles. Since the genetic polymorphisms of the CYP2A6 gene have a major impact on nicotine clearance, its relationships with smoking behavior or the risk of lung cancer have been suggested. The metabolic pathways of the glucuronidation of nicotine, cotinine, and trans-3'-hydroxycotinine in humans would be one of the causal factors for the interindividual differences in nicotine metabolism. This review mainly summarizes recent results from our studies. [Abstract/Link to Full Text]

Nakamura T, Sakaeda T, Takahashi M, Hashimoto K, Gemma N, Moriya Y, Komoto C, Nishiguchi K, Okamura N, Okumura K
Simultaneous determination of single nucleotide polymorphisms of MDR1 genes by electrochemical DNA chip.
Drug Metab Pharmacokinet. 2005 Jun;20(3):219-25.
The on-chip genotyping system ("the electrochemical DNA chip") has been developed as a more cost-effective genotyping system and was applied to MDR1 genotyping in the present study, which is required for wide use in clinical application and for personalized medication based on genotype. The electrochemical DNA chip was optimized and applied to simultaneous genotyping of four MDR1 polymorphisms (T-129C, C1236T, G2677(A,T) and C3435T) using synthetic model oligonucleotide DNA and human genomic DNA. The electrochemical DNA chip successfully gave the T-129C, C1236T, G2677(A,T) and C3435T genotypes, which were completely consistent with those determined by direct sequencing. In conclusion, the electrochemical DNA chip is useful for simultaneous determination of some genotypes and haplotypes, and efficient genotyping using this system can support future genotype-phenotype studies at a large scale. [Abstract/Link to Full Text]

Kaji H, Kume T
Identification of human UDP-glucuronosyltransferase isoform(s) responsible for the glucuronidation of 2-(4-chlorophenyl)- 5-(2-furyl)-4-oxazoleacetic acid (TA-1801A).
Drug Metab Pharmacokinet. 2005 Jun;20(3):212-8.
We characterized the hepatic and intestinal UDP-glucuronosyltransferase (UGT) isoform(s) responsible for the glucuronidation of 2-(4-chlorophenyl)-5-(2-furyl)-4-oxazoleacetic acid (TA-1801A) in humans through several in vitro mechanistic studies. Assessment of a panel of recombinant UGT isoforms revealed the TA-1801A glucuronosyltransferase activity of UGT1A1, UGT1A3, UGT1A7, UGT1A9, and UGT2B7. Kinetic analyses of the TA-1801A glucuronidation by recombinant UGT1A1, UGT1A3, UGT1A9, and UGT2B7 showed that the K(m) value for UGT2B7 was apparently consistent with those in human liver and jejunum microsomes. The TA-1801A glucuronosyltransferase activity in human liver microsomes was inhibited by bilirubin (typical substrate for UGT1A1), propofol (typical substrate for UGT1A9), diclofenac (substrate for UGT1A9 and UGT2B7), and genistein (substrate for UGT1A1, UGT1A3, and UGT1A9). The inhibition by bilirubin, propofol, and diclofenac of the TA-1801A glucuronidation was less pronounced in jejunum microsomes than liver microsomes, suggesting that the contribution of UGT1A1, UGT1A9, and UGT2B7 to the TA-1801A glucuronidation is smaller in the intestine than the liver. In contrast, genistein strongly inhibited the TA-1801A glucuronosyltransferase activity in both human liver and jejunum microsomes. These results suggest that the glucuronidation of TA-1801A is mainly catalyzed by UGT1A1, UGT1A9, and UGT2B7 in the liver, and by UGT1A1, UGT1A3, and UGT2B7 in the intestine in humans. [Abstract/Link to Full Text]

Kaji H, Kume T
Glucuronidation of 2-(4-chlorophenyl)-5-(2-furyl)-4-oxazoleacetic acid (TA-1801A) in humans: species differences in liver and intestinal microsomes.
Drug Metab Pharmacokinet. 2005 Jun;20(3):206-11.
The metabolism of ethyl 2-(4-chlorophenyl)-5-(2-furyl)-4-oxazoleacetate (TA-1801), a potent hypolipidemic agent, was studied in humans after oral administration and compared with that found in rats, rabbits, and dogs previously. Hydrolysis of the ethyl ester to produce metabolite M1 (TA-1801 active form; TA-1801A) is the first metabolic step and the subsequent biotransformation includes the glucuronidation to form the metabolite M4 and the oxidation to form the metabolites M2 and M3. The metabolism of TA-1801 in humans was qualitatively similar to that in the experimental animals studied, although species differences were seen in the amount of metabolites. M4, the glucuronide of TA-1801A was the most abundant metabolite in human urine (24.3% of the dose). In vitro studies using human liver and jejunum microsomes indicated that the TA-1801A glucuronosyltransferase activity in human jejunum microsomes was 2-fold higher than that in liver microsomes. With regard to the interspecies differences in the TA-1801A glucuronosyltransferase activities, the intrinsic clearance for the TA-1801A glucuronidation in liver microsomes was in the following order: rabbit>monkey>human=rat=dog. In jejunum microsomes, the intrinsic clearance for the TA-1801A glucuronidation was in the following order: human>monkey>rabbit>rat=dog. These results suggest that the species differences in the intestinal TA-1801A glucuronidation contribute to the species differences in the excretion rate of TA-1801A glucuronide into the urine. [Abstract/Link to Full Text]

Urakami Y, Kimura N, Okuda M, Masuda S, Katsura T, Inui K
Transcellular transport of creatinine in renal tubular epithelial cell line LLC-PK1.
Drug Metab Pharmacokinet. 2005 Jun;20(3):200-5.
BACKGROUND/AIM: Creatinine is excreted into urine via tubular secretion in addition to glomerular filtration. In the present study, characteristics of the creatinine transport in renal epithelial cells were investigated. METHODS: The transcellular transport and accumulation of [14C]creatinine and [14C]tetraethylammonium (TEA) were assessed using LLC-PK1 cell monolayers cultured on porous membrane filters. RESULTS: [14C]Creatinine was transported directionally from the basolateral to apical side of LLC-PK1 cell monolayers. Basolateral uptake of [14C]creatinine was dependent on membrane potential, and was saturable with apparent K(m) and V(max) values of 13.2+/-2.8 mM and 13.1+/-3.1 nmol/mg protein/5 min, respectively. Concomitant administration of organic cations (1 mM) such as cimetidine, quinidine and trimethoprim inhibited both the transcellular transport and accumulation of [14C]creatinine. Furthermore, apical excretion of [14C]creatinine was not dependent on acidification of the apical medium. CONCLUSIONS: Creatinine was subjected to directional transport across renal epithelial cells from the basolateral to apical side. The organic cation transporter should be involved in the basolateral uptake of creatinine. [Abstract/Link to Full Text]

Matsui-Sakata A, Ohtani H, Sawada Y
Pharmacokinetic-pharmacodynamic analysis of antipsychotics-induced extrapyramidal symptoms based on receptor occupancy theory incorporating endogenous dopamine release.
Drug Metab Pharmacokinet. 2005 Jun;20(3):187-99.
We aimed to analyze the risks of extrapyramidal symptoms (EPS) induced by typical and atypical antipsychotic drugs using a common pharmacokinetic-pharmacodynamic (PK-PD) model based on the receptor occupancy. We collected the data for EPS induced by atypical antipsychotics, risperidone, olanzapine and quetiapine, and a typical antipsychotic, haloperidol from literature and analyzed the following five indices of EPS, the ratio of patients obliged to take anticholinergic medication, the occurrence rates of plural extrapyramidal symptoms (more than one of tremor, dystonia, hypokinesia, akathisia, extrapyramidal syndrome, etc.), parkinsonism, akathisia, and extrapyramidal syndrome. We tested two models, i.e., a model incorporating endogenous dopamine release owing to 5-HT2A receptor inhibition and a model not considering the endogenous dopamine release, and used them to examine the relationship between the D2 receptor occupancy of endogenous dopamine and the extent of drug-induced EPS. The model incorporating endogenous dopamine release better described the relationship between the mean D2 receptor occupancy of endogenous dopamine and the extent of EPS than the other model, as assessed by the final sum of squares of residuals (final SS) and Akaike's Information Criteria (AIC). Furthermore, the former model could appropriately predict the risks of EPS induced by two other atypical antipsychotics, clozapine and ziprasidone, which were not incorporated into the model development. The developed model incorporating endogenous dopamine release owing to 5-HT2A receptor inhibition may be useful for the prediction of antipsychotics-induced EPS. [Abstract/Link to Full Text]

Fukumoto K, Tanemura M, Tsuchishita Y, Kusumoto M, Matsumoto K, Kamakura S, Ueno K
Effect of protein binding of pilsicainide on the pharmacokinetics.
Drug Metab Pharmacokinet. 2005 Jun;20(3):183-6.
To evaluate the effect of protein binding of pilsicainide on its clearance and the contribution of protein binding to optimized pilsicainide therapy, clinical laboratory and pharmacokinetic data were studied in 160 Japanese inpatients (Study 1) and 18 Japanese inpatients (Study 2). To determine the relation between protein concentration and the protein binding ratio of pilsicainide in vitro, the effect of human alpha1-acid glycoprotein (AAG) and human albumin on the binding ratio was studied. The mean ratio of serum pilsicainide concentration to dose per body weight (C/D) increased with increases in the C-reactive protein (CRP) concentration in Study 1. The AAG level increased with increases in the CRP concentration and the binding ratio increases in the AAG concentration in the Study 2. The binding ratios increased with increased AAG and albumin concentrations; the AAG concentration relative to the ratio was particularly large in vitro study. These results suggest C/D is increased in patients with high CRP levels because of binding of pilsicainide to protein, resulting decreased clearance. [Abstract/Link to Full Text]

Takashima T, Murase S, Iwasaki K, Shimada K
Evaluation of dextromethorphan metabolism using hepatocytes from CYP2D6 poor and extensive metabolizers.
Drug Metab Pharmacokinet. 2005 Jun;20(3):177-82.
It is important to estimate the defective metabolism caused by genetic polymorphism of drug metabolizing enzymes before the clinical stage. We evaluated the utility of cryopreserved human hepatocytes of CYP2D6 poor metabolizer (PM) for the estimation of the metabolism in PM using dextromethorphan (DEX) as the probe drug for CYP2D6 substrate. The results of low formations of dextrorphan (DXO) and 3-hydroxymorphinan (3-HM) in CYP2D6 PM hepatocytes incubated with dextromethorphan reflected the clinical data. Formation of 3-methoxymorphinan (3-MEM) normalized by CYP3A4/5 activity in the PM hepatocytes reached about 2.8-fold higher than that in CYP2D6 extensive metabolizer (EM) hepatocytes, which clearly showed the compensatory metabolic pathway of O-demethylation catalyzed by CYP2D6 as seen in clinical study. On the contrary, in the condition of the EM hepatocytes with CYP2D6 inhibitors, the enhancement of 3-MEM formation was not observed. In phase II reaction, the glucuronide formation rate of DXO in the PM hepatocytes was lower than that in the EM hepatocytes, which was consistent with clinical data of DXO-glucuronide (DXO-glu) concentration. These results would suggest that CYP2D6 PM hepatocytes could be a good in vitro tool for estimating CYP2D6 PM pharmacokinetics. [Abstract/Link to Full Text]

Kose N, Yamamoto K, Sai Y, Isawa M, Suwa T, Nakashima E
Prediction of theophylline clearance in CCl4-treated rats using in vivo CYP1A2 and CYP3A2 contents assessed with the PKCYP test.
Drug Metab Pharmacokinet. 2005 Jun;20(3):168-76.
We previously established a method to predict the drug metabolism capacity of injured liver based on pharmacokinetic estimation of the amount of cytochrome P450 (CYP) in vivo (PKCYP test), by introducing the apparent liver-to-blood free concentration gradient in vivo (qg) as a parameter. Here we show that the amount of CYP3A2 in CCl(4)-treated rats can be estimated appropriately by applying the PKCYP test using midazolam (MDZ) as a probe, assuming that the qg value in control rats does not change. We applied the results to predict the clearance of theophylline as a model drug with a physiologically based pharmacokinetic model. Male Sprague-Dawley rats were pretreated with CCl4, and the amount of CYP (A-CYP(vivo)) was quantified by Western blotting. The qg value of MDZ was determined in control rats and used to estimate the amounts of CYP3A2 in CCl4-treated rats; the result agreed well with the observed values. The qg value of CYP3A2 estimated with MDZ as a probe was used together with our previously reported value for CYP1A2 (theophylline metabolism in the liver is known to be almost entirely mediated by CYP3A2 and CYP1A2) to predict the total body clearance (CL(tot)) of theophylline in CCl4-treated rats. The predicted CL(tot) was about one-third of the observed value, which was considered acceptable. The time-course of theophylline concentration in serum simulated with a physiologically-based pharmacokinetic model agreed well with the observed values. Thus, the PKCYP test using MDZ as a probe can be used to predict the amount of CYP3A2 and the CL(tot) of theophylline in CCl4-treated rats. [Abstract/Link to Full Text]

Furuta T, Shirai N, Sugimoto M, Nakamura A, Hishida A, Ishizaki T
Influence of CYP2C19 pharmacogenetic polymorphism on proton pump inhibitor-based therapies.
Drug Metab Pharmacokinet. 2005 Jun;20(3):153-67.
Proton pump inhibitors (PPIs), such as omeprazole, lansoprazole, rabeprazole, esomeprazole, and pantoprazole, are mainly metabolized by CYP2C19 in the liver. There are genetically determined differences in the activity of this enzyme. The genotypes of CYP2C19 are classified into the three groups, rapid extensive metabolizer (RM), intermediate metabolizer (IM), and poor metabolizer (PM). The pharmacokinetics and pharmacodynamics of PPIs depend on CYP2C19 genotype status. Plasma PPI levels and intragastric pHs during PPI treatment in the RM group are lowest, those in the IM group come next, and those in the PM group are highest of the three groups. These CYP2C19 genotype-dependent differences in pharmacokinetics and pharmacodynamics of PPIs influence the cure rates for the gastro-esophageal reflux disease and H. pylori infection by PPI-based therapies. For the better PPI-based treatment, doses and dosing schemes of PPIs should be optimized based on CYP2C19 genotype status. [Abstract/Link to Full Text]

Saeki M, Saito Y, Jinno H, Sai K, Hachisuka A, Kaniwa N, Ozawa S, Kawamoto M, Kamatani N, Shirao K, Minami H, Ohtsu A, Yoshida T, Saijo N, Komamura K, Kotake T, Morishita H, Kamakura S, Kitakaze M, Tomoike H, Sawada J
Genetic variations and haplotypes of UGT1A4 in a Japanese population.
Drug Metab Pharmacokinet. 2005 Apr;20(2):144-51.
Nineteen genetic variations, including 11 novel ones, were found in exon 1 and its flanking region of the UDP-glucuronosyltransferase (UGT) 1A4 gene from 256 Japanese subjects, consisting of 60 healthy volunteers, 88 cancer patients and 108 arrhythmic patients. These variations include -217T>G and -36G>A in the 5'-flanking region, 30G>A (P10P), 127delA (43fsX22; frame-shift from codon 43 resulting in the termination at the 22nd codon, codon 65), 175delG (59fsX6), 271C>T (R91C), 325A>G (R109G), and 357T>C (N119N) in exon 1, and IVS1+1G>T, IVS1+98A>G and IVS1+101G>T in the following intron. Among them, 127delA and 175delG can confer early termination of translation, resulting in an immature protein that probably lacks enzymatic activity. Variation IVS1+1G>T is located at a splice donor site and thus may lead to aberrant splicing. Since we did not find any significant differences in the frequencies of all the variations among the three subject groups, the data were analyzed as one group. The allele frequencies of the novel variations were 0.006 for IVS1+101G>T, 0.004 for 30G>A (P10P) and 357T>C (N119N), and 0.002 for the 8 other variations. In addition, the two known nonsynonymous single nucleotide polymorphisms (SNPs), 31C>T (R11W) and 142T>G (L48V), were found at 0.012 and 0.129 frequencies, respectively. The SNP 70C>A (P24T), mostly linked with 142T>G (L48V) in German Caucasians, was not detected in this study. Sixteen haplotypes were identified or inferred, and some haplotypes were confirmed by cloning and sequencing. It was shown that most of 142T>G (L48V) was linked with -219C>T, -163G>A, 448T>C (L150L), 804G>A (P268P), and IVS1+43C>T, comprising haplotype *3a; haplotype *4a harbors 31C>T (R11W); 127delA (43fsX22) and 142T>G (L48V) were linked (haplotype *5a); 175delG (59fsX6) was linked with 325A>G (R109G) (*6a haplotype); and -219C>T, -163G>A, 142T>G (L48V), 271C>T (R91C), 448T>C (L150L), 804G>A (P268P), and IVS1+43C>T comprised haplotype *7a. Our results provide fundamental and useful information for genotyping UGT1A4 in the Japanese and probably Asian populations. [Abstract/Link to Full Text]

Recent Articles in Journal of Pharmacological Sciences

Chen J, Shen H, Nagasawa Y, Mitsui K, Tsurugi K, Hashimoto K
Pravastatin inhibits arrhythmias induced by coronary artery ischemia in anesthetized rats.
J Pharmacol Sci. 2007 Mar;103(3):317-22.
We have reported that chronically administered pravastatin prevented coronary artery reperfusion-induced lethal ventricular fibrillation (VF) in anesthetized rats without lowering the serum cholesterol level. The present study was undertaken to evaluate whether pravastatin prevents ischemia-induced lethal VF, simultaneously examining myeloperoxidase (MPO) activity in ischemic myocardial tissues. Anesthetized rats were subjected to 30-min ischemia and 60-min reperfusion after chronic administration of pravastatin (0.02, 0.2, and 2 mg/kg), fluvastatin (2 and 4 mg/kg), or vehicle for 22 days, orally, once daily. ECG and blood pressure were continually recorded, and MPO was measured by a spectrophotometer. Pravastatin and fluvastatin significantly (P<0.05) decreased MPO activities, but only pravastatin decreased the incidence of ischemia-induced lethal VF. Both statins had no significant effects on body weight, blood pressure, heart rate, and QT interval as we reported earlier. Our results prove further that pravastatin has benefits to decrease cardiovascular mortality beyond its cholesterol-lowering effect. Pravastatin is more potent than fluvastatin in prevention of arrhythmias. A decrease in the neutrophil infiltration may be partly involved in the inhibitory effect of pravastatin on the ischemia-induced VF. [Abstract/Link to Full Text]

Matsuya T, Takuma K, Sato K, Asai M, Murakami Y, Miyoshi S, Noda A, Nagai T, Mizoguchi H, Nishimura S, Yamada K
Synergistic effects of adenosine A2A antagonist and L-DOPA on rotational behaviors in 6-hydroxydopamine-induced hemi-Parkinsonian mouse model.
J Pharmacol Sci. 2007 Mar;103(3):329-32.
In this study, we examined the combination effects of L-DOPA and adenosine receptor antagonists on rotational behaviors in a hemi-Parkinsonian mouse model induced by unilateral 6-hydroxydopamine (6-OHDA) injection. The adenosine A(2A) antagonist SCH-58261, but not the A(1)-receptor antagonist 8-cyclopentyl-1,3-dipropylxanthine or A(2B)-receptor antagonist alloxazine, synergistically potentiated the L-DOPA-induced rotational behaviors in the 6-OHDA-lesioned mice. In addtion, the 6-OHDA-induced lesions of the dopaminergic system did not affect the in vivo binding of an adenosine A(2A)-receptor tracer [(11)C]SCH-442416 in the striatatum. These findings suggest that adenosine A(2A) antagonists are extremely useful for pharmacotherapy of L-DOPA in Parkinson's disease patients. [Abstract/Link to Full Text]

Yamaguchi Y, Nasu F, Harada A, Kunitomo M
Oxidants in the gas phase of cigarette smoke pass through the lung alveolar wall and raise systemic oxidative stress.
J Pharmacol Sci. 2007 Mar;103(3):275-82.
Cigarette smoking-induced oxidative stress plays a key role in the pathogenesis of atherosclerosis in smokers. Aqueous cigarette smoke extract (CSE) contains stable oxidants, peroxynitrite-like reactants, which have the ability to oxidize and nitrate low-density lipoprotein (LDL). We examined whether oxidants in CSE can penetrate into the blood through the lung alveolar wall and cause oxidative vascular injury. The oxidants in CSE and sodium peroxynitrite could easily pass through the reconstituted basement membrane. When CSE or sodium peroxynitrite solution was infused into the alveolar air space of an isolated rat lung mounted in tyrosine solution, CSE gradually increased the 3-nitrotyrosine levels in the external tyrosine solution while sodium peroxynitrite caused a rapid increase. CSE did not activate the rat alveolar macrophages. When rats were acutely exposed to the gas phase of cigarette smoke from which tar and nicotine had been removed, both serum levels of 3-nitrotyrosine and 8-hydroxy-2'-deoxyguanine, oxidative stress markers, rapidly increased. Our results demonstrate that relatively stable oxidants in CSE can pass through the pulmonary alveolar wall into the blood and induce systemic oxidative stress, which most likely facilitates oxidative modification of LDL and endothelial dysfunction, explaining early key events in the development of atherosclerosis. [Abstract/Link to Full Text]

Hinata M, Matsuoka I, Iwamoto T, Watanabe Y, Kimura J
Mechanism of Na+/Ca2+ exchanger activation by hydrogen peroxide in guinea-pig ventricular myocytes.
J Pharmacol Sci. 2007 Mar;103(3):283-92.
Using the whole-cell voltage clamp, we examined the mechanism of activation of the Na(+)/Ca(2+) exchanger (NCX) by hydrogen peroxide (H(2)O(2)) in isolated guinea-pig cardiac ventricular myocytes. Exposure to H(2)O(2) increased the NCX current. The effect was inhibited by cariporide, an inhibitor of the Na(+)/H(+) exchanger (NHE), suggesting that there are NHE-dependent and -independent pathways in the effect of H(2)O(2) on NCX. In addition, both pathways were blocked by edaravone, a hydroxyl radical (*OH) scavenger; pertussis toxin, a Galpha(i/o) protein inhibitor; and U0126, an inhibitor of mitogen-activated protein kinase/extracellular signal-regulated kinase kinase (MEK). On the other hand, wortmannin, a phosphatidylinositol 3-kinase (PI3K) inhibitor, inhibited only the NHE-dependent pathway, while PP2, a Src family protein tyrosine kinase inhibitor, inhibited only the NHE-independent pathway. Taken together, our data suggest that H(2)O(2) increases the NCX current via two signal transduction pathways. The common pathway is the conversion of H(2)O(2) to *OH, which activates Galpha(i/o) protein and a mitogen-activated protein (MAP) kinase signaling pathway. Then, one pathway activates NHE with a PI3K-dependent mechanism and indirectly increases the NCX current. Another pathway involves activation of a Src family tyrosine kinase. [Abstract/Link to Full Text]

Nishioku T, Takata F, Yamauchi A, Sumi N, Yamamoto I, Fujino A, Naito M, Tsuruo T, Shuto H, Kataoka Y
Protective action of indapamide, a thiazide-like diuretic, on ischemia-induced injury and barrier dysfunction in mouse brain microvascular endothelial cells.
J Pharmacol Sci. 2007 Mar;103(3):323-7.
The aim of the present study was to elucidate the effects of indapamide on ischemic damage to the blood-brain barrier (BBB) in vitro. The ischemia/reperfusion conditions employed here significantly decreased the viability of mouse brain capillary endothelial (MBEC4) cells, an effect ameliorated by indapamide. Ischemia increased the permeability of MBEC4 cells to two cellular transport markers, sodium fluorescein and Evan's blue-albumin. Indapamide reduced the ischemia-induced hyperpermeability of cells. These results suggest that indapamide may have a protective role against ischemia-induced injury and dysfunction of the BBB. [Abstract/Link to Full Text]

Jadhav U, Ezhilarasan R, Vaughn SF, Berhow MA, Mohanam S
Dietary isothiocyanate iberin inhibits growth and induces apoptosis in human glioblastoma cells.
J Pharmacol Sci. 2007 Feb;103(2):247-51.
In this study, we evaluated the antiproliferative and proapoptotic effects of the isothiocyanate iberin, a bioactive agent in Brassicaceae species, in human glioblastoma cells. The human glioblastoma cell cultures were treated with different concentrations of iberin and tested for growth inhibition, cytotoxicity, induction of apoptosis, and activation of caspases. Iberin inhibited growth of tumor cells in cell proliferation assays, enhanced cytotoxicity, and induced apoptosis by activation of caspase-3 and caspase-9. Findings from this study could provide a basis for potential usefulness of the diet-derived isothiocyanate iberin as a promising therapeutic micronutrient in the prevention/intervention of brain tumors. [Abstract/Link to Full Text]

Tanaka H, Shimada H, Namekata I, Kawanishi T, Iida-Tanaka N, Shigenobu K
Involvement of the Na+/Ca2+ exchanger in ouabain-induced inotropy and arrhythmogenesis in guinea-pig myocardium as revealed by SEA0400.
J Pharmacol Sci. 2007 Feb;103(2):241-6.
Involvement of the Na+/Ca2+ exchanger in ouabain-induced inotropy and arrhythmogenesis was examined with a specific inhibitor, SEA0400. In right ventricular papillary muscle isolated from guinea-pig ventricle, 1 microM SEA0400, which specifically inhibits the Na+/Ca2+ exchanger by 80%, reduced the ouabain (1 microM)-induced positive inotropy by 40%, but had no effect on the inotropy induced by 100 microM isobutyl methylxantine. SEA0400 significantly inhibited the contracture induced by low Na+ solution. In HEK293 cells expressing the Na+/Ca2+ exchanger, 1 microM ouabain induced an increase in intracellular Ca2+, which was inhibited by SEA0400. The arrhythmic contractions induced by 3 microM ouabain were significantly reduced by SEA0400. These results provide pharmacological evidence that the Na+/Ca2+ exchanger is involved in ouabain-induced inotropy and arrhythmogenesis. [Abstract/Link to Full Text]

Sam TS, Ngan MP, Riendeau D, Robichaud A, Rudd JA
Action of cyclooxygenase inhibitors and a leukotriene biosynthesis inhibitor on cisplatin-induced acute and delayed emesis in the ferret.
J Pharmacol Sci. 2007 Feb;103(2):189-200.
Cisplatin at 5 mg/kg, i.p. induced an acute (day 1) and delayed (days 2 and 3) emetic response in the ferret that was used to investigate the anti-emetic activity of the non-selective cyclooxygenase inhibitor indomethacin (3 - 30 mg/kg, i.p., three times per day) and two cyclooxygenase-2 inhibitors, DFU [5,5-dimethyl-3-(3-fluorophenyl)-4-(4-methylsulphonyl)phenyl-2(5H)-furanone; 1 - 10 mg/kg, i.p. administered at 40 and 48 h] and L-745,337 [5-methanesulphonamido-6-(2,4-diflurothiophenyl)-1-indanone; 10 mg/kg, i.p., administered at 40 and 48 h]. Only indomethacin potentiated significantly cisplatin-induced retching + vomiting (P<0.05); DFU antagonized delayed emesis (P<0.05) but the action was not dose-related and L-745,337 was inactive (P>0.05). However, indomethacin alone (30 mg/kg) also induced emesis (P<0.05). The leukotriene biosynthesis inhibitor, MK-886 {3-[1-(p-chlorobenzyl)-5-(isopropyl)-3-tert-butylthioindol-2-yl]-2,2-dimethylpropanoic acid; 1 - 10 mg/kg, i.p., three times per day} had no action to modify cisplatin-induced emesis (P>0.05). The combination treatment of indomethacin (10 mg/kg, i.p., three times per day) with MK-886 (10 mg/kg, i.p., three times per day) did not antagonize cisplatin-induced acute delayed retching + vomiting and had a different profile compared to the action of dexamethasone (1 mg/kg, i.p., three times per day; P<0.05). Inhibition of the cyclooxygenase and lipoxygenase pathways does not account for the anti-emetic of dexamethasone. [Abstract/Link to Full Text]

Shibata S
Circadian rhythms in the CNS and peripheral clock disorders: preface.
J Pharmacol Sci. 2007 Feb;103(2):133. [Abstract/Link to Full Text]

Maemura K, Takeda N, Nagai R
Circadian rhythms in the CNS and peripheral clock disorders: role of the biological clock in cardiovascular diseases.
J Pharmacol Sci. 2007 Feb;103(2):134-8.
The cardiovascular diseases are closely related to circadian rhythm, which is under the control of the biological clock. Clock genes show circadian oscillation not only in the suprachiasmatic nucleus but also in peripheral tissues, suggesting the existence of the peripheral clock. We previously demonstrated that plasminogen activator inhibitor-1 (PAI-1) might be an output gene of the peripheral clock. To further elucidate the functional relevance of the peripheral clock in the cardiovascular system, we screened target genes of the peripheral clock by cDNA microarray analysis. A total of 29 genes including transcription factor, growth factors, and membrane receptors were upregulated by CLOCK/BMAL and showed circadian oscillation. These results suggest that cardiovascular systems have their own peripheral clocks, and at least in part, they may regulate the circadian oscillation of cardiovascular function directly. These results potentially provide a molecular basis for the circadian variation of cardiovascular function and novel strategies for the prevention and treatment of cardiovascular diseases. [Abstract/Link to Full Text]

Kudo T, Horikawa K, Shibata S
Circadian rhythms in the CNS and peripheral clock disorders: the circadian clock and hyperlipidemia.
J Pharmacol Sci. 2007 Feb;103(2):139-43.
A circadian clock controls various physiological and behavioral rhythms. In mammals, a master circadian clock exists in the suprachiasmatic nucleus of the hypothalamus, and slave oscillators can be found in most tissues. These circadian oscillations are controlled by "clock genes". The negative feedback loop is thought to function as a molecular mechanism of the circadian clock. It is plausible that clock genes may control lipid metabolism through so-called clock-controlled genes and that lipid metabolism-related clock-controlled genes may play important roles in the circadian change of lipid metabolism. Recently research has focused on the relationships between the clock system and lipid metabolism. In this review, we discuss the following items: 1) circadian clock system, 2) effect of the diet on clock gene expression, 3) effect of clock mutation on lipid metabolism, and 4) effect of streptozotocin-induced diabetes and ob mutation on clock gene expression and lipid metabolism. In this review we have summarized how the circadian clock affects lipid metabolism through the expression of lipid metabolism-related clock-controlled genes and at the same time discussed how abnormal metabolism of lipid affects the expression of clock genes. Further experiments are needed to elucidate the detailed mechanism of interaction between clock genes and lipid metabolisms. [Abstract/Link to Full Text]

Burioka N, Fukuoka Y, Takata M, Endo M, Miyata M, Chikumi H, Tomita K, Kodani M, Touge H, Takeda K, Sumikawa T, Yamaguchi K, Ueda Y, Nakazaki H, Suyama H, Yamasaki A, Sano H, Igishi T, Shimizu E
Circadian rhythms in the CNS and peripheral clock disorders: function of clock genes: influence of medication for bronchial asthma on circadian gene.
J Pharmacol Sci. 2007 Feb;103(2):144-9.
Bronchial asthma is a chronic inflammatory disorder of the airways, in which inflammation causes bronchial hyper-responsiveness and flow limitation in the presence of various stimuli. Pulmonary function in asthmatic patients frequently deteriorates between midnight and early morning, which has suggested a role for chronotherapy. Although relationships between bronchial asthma and the function of clock genes remain unclear, some medications given for asthma such as glucocorticoids or beta(2)-adrenoceptor agonists may influence clock genes in vivo. In our studies of clock gene mRNA expressions in human bronchial epithelial cells in vitro and peripheral blood cells in vivo, we demonstrated that glucocorticoid or beta(2)-adrenoceptor agonist treatment strongly induced human Per1 mRNA expression both in vitro and in vivo. Human peripheral blood cells provide a useful indication of peripheral clock gene mRNA expression in vivo. [Abstract/Link to Full Text]

Ebisawa T
Circadian rhythms in the CNS and peripheral clock disorders: human sleep disorders and clock genes.
J Pharmacol Sci. 2007 Feb;103(2):150-4.
Genetic analyses of circadian rhythm sleep disorders (CRSD), such as familial advanced sleep phase syndrome (ASPS) and delayed sleep phase syndrome (DSPS), and morningness-eveningness revealed the relationship between variations in clock genes and diurnal change in human behaviors. Variations such as T3111C in the Clock gene are reportedly associated with morningness-eveningness. Two of the pedigrees of familial ASPS (FASPS) are caused by mutations in clock genes: the S662G mutation in the Per2 gene or the T44A mutation in the casein kinase 1 delta (CK1delta) gene, although these mutations are not found in other pedigrees of FASPS. As for DSPS, a missense variation in the Per3 gene is identified as a risk factor, while the one in the CK1epsilon gene is thought to be protective. These findings suggest that further, as yet unidentified, gene variations are involved in human circadian activity. Many of the CRSD-relevant variations reported to date seem to affect the phosphorylation status of the clock proteins. A recent study using mathematical models of circadian rhythm generation has provided a new insight into the role of phosphorylation in the molecular mechanisms of these disorders. [Abstract/Link to Full Text]

Ohdo S
Circadian rhythms in the CNS and peripheral clock disorders: chronopharmacological findings on antitumor drugs.
J Pharmacol Sci. 2007 Feb;103(2):155-8.
The effectiveness and toxicity of antitumor drugs vary depending on dosing time associated with the 24-h rhythms of biochemical, physiological, and behavioral processes under the control of the circadian clock. Such chronopharmacological phenomena are influenced by not only the pharmacokinetics but also pharmacodynamics of medications. For example, the antitumor effect and/or toxicity of irinotecan hydrochloride, interferon, and antiangiogenic agents vary depending on the dosing time associated with the 24-h rhythm of their target enzyme, receptor, protein, and pharmacokinetics. Many of them are controlled by clock genes. Chronotherapy is especially relevant when the risk and/or intensity of the symptoms of disease vary predictably over time. In a randomized multicenter trial involving patients with previously untreated metastases from colorectal cancer, the chronomodulated infusion of oxaliplatin, fluorouracil (5-FU), and folinic acid is compared with a constant-rate infusion method. Side effects such as stomatitis, peripheral sensory neuropathy are lower and the objective response is higher in the chronotherapy as compared with the fixed-rate infusion. The merit of chronomodulated infusion is supported by the 24-h rhythm of DNA synthesis and the activity of dehydropyrimidine dehydrogenase, which brings about the intracellular catabolism of 5-FU. Although interferon (IFN) also alters the clock function, the disruptive effect of IFN on clock function can be overcome by devising a dosing regimen that minimizes adverse drug effects on clock function. Thus one approach to increasing the efficiency of pharmacotherapy is the administration of drugs at times at which they are most effective and/or best tolerated. [Abstract/Link to Full Text]

Kamei J, Hirose N, Oka T, Miyata S, Saitoh A, Yamada M
Effects of methylphenidate on the hyperemotional behavior in olfactory bulbectomized mice by using the hole-board test.
J Pharmacol Sci. 2007 Feb;103(2):175-80.
The most consistent behavioral changes caused by olfactory bulbectomy are hyperemotional responses such as hyperactivity in a novel environment. However, the changes in the emotional behavior of mice after undergoing olfactory bulbectomy have not yet been described in detail. The effects of methylphenidate on the hyperemotional behavior of olfactory bulbectomized (OBX) mice were examined by using the hole-board test. Mice (4-week-old) were subjected to olfactory bulbectomy, and the behavioral test was performed 2 weeks after surgery. OBX mice showed a significant increase in the number of head-dips as compared to the sham-operated mice. This increase was significantly decreased after treatment with methylphenidate (10 microg/kg, s.c.). The norepinephrine (NE) turnover ratio in the frontal cortex in OBX mice was significantly less than that in the sham-operated mice. However, the decreased NE ratio in OBX mice normalized after treatment with methylphenidate. Our results suggest that the increased head-dipping behavior in OBX mice might reflect an impulsive-like behavior. In addition, we proposed that the improvement in the noradrenergic abnormalities in the frontal cortex due to methylphenidate treatment may play a key role in the improvement of impulsive-like behaviors observed in OBX mice. [Abstract/Link to Full Text]

Wang K, Takahara A, Nakamura Y, Aonuma K, Matsumoto M, Sugiyama A
In vivo electropharmacological effects of amiodarone and candesartan on atria of chronic atrioventricular block dogs.
J Pharmacol Sci. 2007 Feb;103(2):207-13.
Electropharmacological effects of chronically administered amiodarone and candesartan on atria that had been remodeled against congestive heart failure were assessed using dogs (about 10 kg in weight) with chronic atrioventricular block. Amiodarone was administered orally in a dose of 200 mg/body per day for the initial 7 days followed by 100 mg for the following 21 days (n = 7). Candesartan was administered in a dose of 12 mg/body per day for 28 days (n = 7). All animals survived the 4-week experimental period, indicating the lack of risks for inducing cardiohemodynamic collapse or torsade de pointes by these drugs. The plasma amiodarone concentration was 353 ng/ml at 4 weeks of treatment. Before candesartan treatment (control), intravenous administration of 30 ng/kg of angiotensin II increased the mean blood pressure by 18 mmHg, which was significantly decreased to 1 mmHg by 4 weeks of treatment. Amiodarone prolonged the atrial effective refractory period without affecting inter-atrial conduction time and decreased the duration of the burst pacing-induced atrial fibrillation, whereas candesartan hardly affected these variables. These results indicate that amiodarone should become a pragmatic pharmacological strategy against atrial fibrillation in patients with chronically compensated heart failure and suggest that a much higher dose of candesartan may be needed to exert its efficacy in this model. [Abstract/Link to Full Text]

Kawasumi H, Satoh N, Kitada Y
Caldaret, an intracellular Ca2+ handling modulator, limits infarct size of reperfused canine heart.
J Pharmacol Sci. 2007 Feb;103(2):222-33.
The cardioprotective effect of caldaret, a novel intracellular Ca(2+) handling modulator that acts through reverse-mode Na(+)/Ca(2+) exchanger inhibition and potential sarcoplasmic reticulum (SR) Ca(2+) uptake enhancement, against reperfusion injury was investigated. We employed a canine model of myocardial infarction induced by 90-min occlusion of left circumflex (LCX) coronary artery followed by 4 h of reperfusion. Intravenously infused caldaret (3 or 30 microg/kg per hour) for 30 min at LCX-reperfusion markedly reduced infarct size (by 51.3% or 71.9%, respectively). This cardioprotection was accompanied by an acceleration of left ventricular (LV) contraction and relaxation during reperfusion, but not by an increase in ischemic regional transmural myocardial blood flow (TMBF) or endocardial/epicardial blood flow ratio (Endo/Epi ratio) or a reduction in double-product throughout the protocol. Diltiazem (2000 microg/kg per hour) also reduced infarct size (by 36.1%), but unlike caldaret, was accompanied by the significant increase in Endo/Epi ratio in the ischemic region and decrease in double-product. There were significant inverse relationships between infarct size and ischemic regional TMBF in all groups. Caldaret, but not diltiazem shifted the regression line downward with a flatter slope. These results suggest that the amelioration of intracellular Ca(2+) handling dysfunction achieved by caldaret leads to cardioprotective effects against reperfusion injury following prolonged ischemia. [Abstract/Link to Full Text]

Miyata T
Pharmacological basis of traditional medicines and health supplements as curatives.
J Pharmacol Sci. 2007 Feb;103(2):127-31.
Traditional Oriental medicines and health supplements have been empirically used to treat various ailments but most of them have not been evaluated objectively to prove their efficacies. We have been investigating the medical benefits of traditional Oriental medicines and health supplements as alternatives and their varied actions and mechanisms by pharmacological approaches. The study on airway inflammation has shown that even a Kampo preparation, Bakumondo-to, has anti-inflammatory, anti-allergic, immunomodulatory, secretory-modulating, and metabolic regulatory actions. All of its actions are based on the restoration of normal molecular and cellular functions through DNA transcriptional regulation. In other previous studies, we showed that a health supplement, royal jelly (RJ) has weak estrogenic activity. RJ competes with 17beta-estradiol for binding to the human estrogen receptors alpha and beta, although it is much weaker than diethylstilbestrol in binding affinity. Treatment of MCF-7 cells with RJ enhances proliferation, and concomitant treatment with tamoxifen blocked this effect. A reporter gene assay showed that RJ enhanced transcription of the luciferase gene through the estrogen-responsive element in MCF-7 cells. Furthermore, subcutaneous injection of RJ restored the expression of vascular endothelial growth factor gene in the uteri of ovariectomized rats. We suggest that the diverse pharmacological functions of RJ can be ascribed, in part, to its estrogenic effects. We hypothesize that traditional medicine, which has multiple actions, may be better than Western medicine of a single component to treat various diseases including "Mibyou" (presymptomatic disease). Our findings provide new ideas about the nature of disorders nd disease-state development that involve complicated mechanisms and will contribute to novel principles to prevent diseases and establish new treatments. Adoption of the means of translational research should provide an objective background for efficacy and stimulate broader application and usage of traditional medicines and health supplements. [Abstract/Link to Full Text]

Kawai H, Suzuki T, Kobayashi T, Ishii-Watabe A, Sakurai H, Ohata H, Honda K, Momose K, Hayakawa T, Kawanishi T
Caspase cascade proceeds rapidly after cytochrome c release from mitochondria in tumor necrosis factor-alpha-induced cell death.
J Pharmacol Sci. 2007 Feb;103(2):159-67.
The caspase activation cascade and mitochondrial changes are major biochemical reactions in the apoptotic cell death machinery. We attempted to clarify the temporal relationship between caspase activation, cytochrome c release, mitochondrial depolarization, and morphological changes that take place during tumor necrosis factor (TNF)-alpha-induced cell death in HeLa cells. These reactions were analyzed at the single-cell level with 0.5 - 1 min resolution by using green fluorescent protein (GFP)-variant-derived probes and chemical probes. Cytochrome c release, caspase activation, and cellular shrinkage were always observed in this order within 10 min in all dying cells. This sequence of events was thus considered a critical pathway of cell death. Mitochondrial depolarization was also observed in all dying cells observed, but frequently occurred after caspase activation and cellular shrinkage. Mitochondrial depolarization is therefore likely to be a reaction that does not induce caspase activation and subsequent cellular shrinkage. Mitochondrial changes are important for apoptotic cell death; moreover, cytochrome c release, and not depolarization, is a key reaction related to cell death. In addition, we also found that the apoptotic pathway proceeds only when cells are exposed to TNF-alpha. These findings suggest that the entire cell death process proceeds rapidly during TNF-alpha exposure. [Abstract/Link to Full Text]

Takeuchi R, Matsumoto H, Okada H, Hori M, Gunji A, Hakozaki K, Akimoto Y, Fujii A
Differences of cell growth and cell cycle regulators induced by basic fibroblast growth factor between nifedipine responders and non-responders.
J Pharmacol Sci. 2007 Feb;103(2):168-74.
Differences of cell proliferation, cell cycle, and G(1)/S transition regulatory proteins of gingival fibroblasts derived from nifedipine-reactive patient (NIFr) and nifedipine-non-reactive patient (NIFn) in the presence of basic fibroblast growth factor (bFGF) were investigated to elucidate the mechanism of gingival overgrowth associated with nifedipine, one of the Ca(2+)-channel blockers. The proliferation rate of NIFr cells in the presence of bFGF significantly increased than NIFn cells. The proportion of NIFr cells that had undergone progression to the S and G(2)/M phases from the G(0)/G(1) phase significantly increased compared to that in NIFn cells. Increases of pRB (Ser807/811), pCDK2 (Thr160), CDK2, and cyclin E protein levels in NIFr cells were greater than those in NIFn cells. The elevations of pRB (Ser780), RB, and cyclin A protein levels in NIFr cells did not differ from those of NIFn cells. The growth of NIFr cells was greater than that of NIFn cells as a result of the active G(1)/S transition of NIFr cells, as assessed by the increments of cyclin E, pCDK2, and pRB (ser807/811) protein in NIFr cells. [Abstract/Link to Full Text]

Takahara A, Nakamura H, Nouchi H, Tamura T, Tanaka T, Shimada H, Tamura M, Tsuruoka N, Takeda K, Tanaka H, Shigenobu K, Hashimoto K, Sugiyama A
Analysis of arrhythmogenic profile in a canine model of chronic atrioventricular block by comparing in vitro effects of the class III antiarrhythmic drug nifekalant on the ventricular action potential indices between normal heart and atrioventricular block heart.
J Pharmacol Sci. 2007 Feb;103(2):181-8.
The chronic atrioventricular block dog is a useful model for predicting the future onset of drug-induced long QT syndrome in clinical practice. To better understand the arrhythmogenic profile of this model, we recorded the action potentials of the isolated ventricular tissues in the presence and absence of the class III antiarrhythmic drug nifekalant. The action potential durations of the Purkinje fiber and free wall of the right ventricle were longer in the chronic atrioventricular block dogs than in the dogs with normal sinus rhythm. Nifekalant in concentrations of 1 and 10 microM prolonged the action potential durations of Purkinje fiber and the free wall in a concentration-dependent manner. The extent of prolongation was greater in the chronic atrioventricular block dogs than in the normal dogs. However, increase of temporal dispersion of ventricular repolarization including early afterdepolarization was not detected by nifekalant in either group of dogs, indicating lack of potential to trigger arrhythmias in vitro. These results suggest that the ventricular repolarization delay in the chronic atrioventricular block model by nifekalant may largely depend on the decreased myocardial repolarization reserve, whereas the trigger for lethal arrhythmia was not generated in the in vitro condition in contrast to the in vivo experiment. [Abstract/Link to Full Text]

Tokunaga S, Takeda Y, Shinomiya K, Hirase M, Kamei C
Effects of some H1-antagonists on the sleep-wake cycle in sleep-disturbed rats.
J Pharmacol Sci. 2007 Feb;103(2):201-6.
The present study was undertaken to investigate the effects of some H(1)-antagonists on the sleep-wake cycle in sleep-disturbed rats in comparison with those of nitrazepam. Electrodes were chronically implanted into the frontal cortex and the dorsal neck muscle of rats for the electroencephalogram (EEG) and electromyogram (EMG), respectively. EEG and EMG were recorded with an electroencephalograph. SleepSign ver. 2.0 was used for EEG and EMG analysis. The total times of waking, non-rapid eye movement (non-REM), and rapid eye movement (REM) sleep were measured from 10:00 to 16:00. Nitrazepam showed a significant decrease in sleep latency, total waking time, and delta activity and an increase in the total non-REM sleep time. A significant decrease in the sleep latency was observed with diphenhydramine, chlorpheniramine, and cyproheptadine. Cyproheptadine also caused a significant decrease in the total waking time and increases in total non-REM sleep time, REM sleep time, slow wave sleep, and delta activity, although no remarkable effects were observed with diphenhydramine and chlorpheniramine. In conclusion, cyproheptadine can be useful as a hypnotic, having not only sleep inducing-effects, but also sleep quantity- and quality-increasing effects. [Abstract/Link to Full Text]

Matsuda K, Nishimura Y, Kurata N, Iwase M, Yasuhara H
Effects of continuous ingestion of herbal teas on intestinal CYP3A in the rat.
J Pharmacol Sci. 2007 Feb;103(2):214-21.
Tenryocha, rooibos, and guava teas are widely consumed as herbal beverages, especially as a therapy against pollen allergy. To investigate the possible herbal tea-drug interaction the effect of continuous ingestion of these teas on cytochrome P450 (CYP) 3A were studied. Rats (n = 6) were allowed free access to either tea (experimental groups) or water (control) for two weeks. Midazolam (MDZ) (20 mg/kg) was orally administered and the serum concentration was determined. The area under the serum concentration-time curve (AUC(0-infinity)) and the maximum serum concentrations (C(max)) of MDZ were reduced by more than 60% after the treatment of tenryocha and rooibos tea (P<0.05). Intestinal MDZ 1'- and 4-hydroxylation activities mediated by CYP3A were increased in tenryocha and rooibos tea-treated group by 50% compared to the control group, although the results were not statistically significant. Furthermore, the Western blot analysis showed that CYP3A content was significantly increased in the intestine after the treatment of these teas (P<0.05). Hepatic MDZ hydroxylation and CYP3A content were slightly increased by these teas. The results suggested that two weeks ingestion of tenryocha and rooibos tea reduced serum concentration of MDZ by the induction of intestinal CYP3A. The possible interaction between tenryocha or rooibos tea and medicines mediated by CYP3A was suggested. [Abstract/Link to Full Text]

Ogura H, Nakanishi-Ueda T, Ueda T, Iwai S, Uchida S, Saito Y, Taguchi Y, Yasuhara H, Armstrong D, Oguchi K, Koide R
Effect of a dihydrobenzofuran derivative on lipid hydroperoxide-induced rabbit corneal neovascularization.
J Pharmacol Sci. 2007 Feb;103(2):234-40.
The aim of this study was to investigate the effect of A-3922, a dihydrobenzofuran derivative, on linoleic acid hydroperoxide (LHP)-induced corneal neovascularization (NV) in a rabbit model. Male New Zealand rabbits received intraperitoneal (i.p.) injections of 10 or 30 mg/kg per day A-3922 or its vehicle as control for 3 days. One day after i.p. injections, LHP was injected with a 30-gauge needle into the corneal stroma of the superior quadrant 4.5-mm below the limbus. Photographs of the vessels were taken for digital analysis with a surgical microscope. Vascular endothelial growth factor (VEGF) was measured using an immunoassay kit, and matrix metalloproteinase (MMP)-9 was measured by gelatin zymography in corneal samples. At 7 days post-LHP injection, the total vessel length was 26.7 +/- 3.8 mm in the control animals (n = 8), 16.1 +/- 0.8 mm in the A-3922 (10 mg/kg)-treated group (n = 5), and 11.4 +/- 2.1 mm in the 30 mg/kg group (n = 8, P<0.01 vs control), respectively. After LHP injection, the content of VEGF and MMP-9 activity were increased in the superior cornea, but these were not influenced by A-3922 treatments. These results indicate that LHP-induced corneal NV is inhibited by treatment with A-3922 and therefore may represent a potential pharmacological intervention for ocular neovascularization disorders. [Abstract/Link to Full Text]

Hatip-Al-Khatib I, Iwasaki K, Egashira N, Ishibashi D, Mishima K, Fujiwara M
Comparison of single- and repeated-ischemia-induced changes in expression of flip and flop splice variants of AMPA receptor subtypes GluR1 and GluR2 in the rats hippocampus CA1 subregion.
J Pharmacol Sci. 2007 Jan;103(1):83-91.
In addition to their role in physiological activities, ionotropic glutamate alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionate receptors (AMPARs) play an important role in neuronal death, especially that following ischemic insults. In this study, we examined the effect of single (SI) and twice repeated (RI)-4-vessel occlusion-ischemia on rat performance in the 8-armed radial maze test. Moreover, the effects of SI and RI on the AMPARs subunits glutamate receptor (GluR) 1 and GluR2 flip and flop variants composition in the CA1 subregion of the hippocampus were investigated using RT-PCR, normalized to glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and expressed as their ratios to the latter. The results showed that SI and RI impaired the maze performance by decreasing correct choices and increasing the error choices, but RI increased error choices to a greater extent than the SI. The SI reduced only GluR1 flip/GAPDH on day 1. The SI did not alter ratios of GluR2 variants to those of GluR1. On the other hand, the RI decreased GluR2 flip and flop variants after 1 and 3 days, respectively, whereas after 7 days, it increased the flip variant of both GluR1 and GluR2. Moreover, the RI reduced ratios of GluR2 variants to those of GluR1. These results reveal the differential effects of the SI and RI on memory and expression of the AMPARs subunits GluR1 and GluR2 and their flip and flop variants in the CA1. [Abstract/Link to Full Text]

Smulders RA, Smith NN, Krauwinkel WJ, Hoon TJ
Pharmacokinetics, safety, and tolerability of solifenacin in patients with renal insufficiency.
J Pharmacol Sci. 2007 Jan;103(1):67-74.
To evaluate the pharmacokinetics, safety, and tolerability of solifenacin in patients with mild, moderate, or severe renal disease, eighteen patients with renal disease and six healthy volunteers received a single oral dose of solifenacin (10 mg). Pharmacokinetic parameters were assessed from blood samples drawn over a 360-h period. Safety and tolerability were also evaluated. Total mean +/- S.D. exposure (ng . h/mL) to solifenacin in healthy individuals (1190 +/- 403) was increased in patients with renal disease (mild: 1784 +/- 792, moderate: 1559 +/- 555, severe: 2530 +/- 700), and elimination half-life (mean +/- S.D. [h]) was prolonged (healthy: 68.2 +/- 27.2, mild: 89.1 +/- 34.5, moderate: 90.6 +/- 27.3, severe: 111 +/- 38.3). A significant correlation was found between creatinine clearance and pharmacokinetic parameters for exposure and apparent oral clearance. No deaths or serious adverse events occurred during the study. Solifenacin 10 mg was well tolerated in patients with renal disease. Solifenacin displays a higher exposure and a prolonged half-life in patients with renal impairment, especially severe. Therefore, while no special cautions are necessary for patients with mild/moderate renal impairment, patients with severe renal impairment should receive no more than 5 mg solifenacin once daily. [Abstract/Link to Full Text]

Li D, Wu LJ, Tashiro S, Onodera S, Ikejima T
Oridonin-induced A431 cell apoptosis partially through blockage of the Ras/Raf/ERK signal pathway.
J Pharmacol Sci. 2007 Jan;103(1):56-66.
We have reported that oridonin, a diterpenoid isolated from the plant Rabdosia rubescens, had apoptosis-inducing activities in many cell lines (e.g., human melanoma A375-S2, human cervical cancer HeLa, human breast adenocarcinoma MCF-7, and murine fibrosarcoma L929). In this study, we further investigated signaling events involved in oridonin-induced apoptosis in human epidermoid carcinoma A431 cells. It was found that the total tyrosine kinase activity was inhibited and the protein expressions of epidermal growth factor receptor (EGFR) and phosphorylated EGFR were decreased in oridonin-induced A431 cell apoptosis. Expression of EGFR downstream effector proteins, Grb2, Ras, Raf-1, and extracellular signal-regulated kinase (ERK), was also downregulated by oridonin. Moreover, the oridonin-induced apoptosis was augmented by the Ras inhibitor manumycin A, Raf-1 inhibitor GW5074, or ERK inhibitor PD98059, suggesting that inactivation of Ras, Raf, or ERK participates in oridonin-induced apoptosis. Taken together, oridonin-induced apoptosis in A431 cells might be through blocking EGFR and its downstream Ras/Raf/ERK signal pathway. [Abstract/Link to Full Text]

Natesan S, Badami S, Dongre SH, Godavarthi A
Antitumor activity and antioxidant status of the methanol extract of Careya arborea bark against Dalton's lymphoma ascites-induced ascitic and solid tumor in mice.
J Pharmacol Sci. 2007 Jan;103(1):12-23.
Based on the ethnomedical use of Careya arborea Roxb bark in the treatment of tumors, the present study was carried out to evaluate the anticancer potentials against Dalton's lymphoma ascites (DLA)-induced ascitic and solid tumors. The methanol extract of its bark given orally to mice at the dose of 250 or 500 mg/kg body weight for 10 days caused significant reduction in percent increase in body weight, packed cell volume, and viable tumor cell count when compared to the mice of the DLA control group. Restoration of hematological and biochemical parameters towards normal was also observed. Histological observations of liver and kidney also indicated repair of tissue damage caused by tumor inoculation. The extract at the dose of 5 or 25 mg/kg body weight given i.p. daily for 14 days significantly reduced the solid tumor volume induced by DLA cells. [Abstract/Link to Full Text]

Godfraind T
Obituary--Dr. Setsuro Ebashi.
J Pharmacol Sci. 2007 Jan;103(1):1-3. [Abstract/Link to Full Text]

Mori A, Saito M, Sakamoto K, Nakahara T, Ishii K
Intravenously administered vasodilatory prostaglandins increase retinal and choroidal blood flow in rats.
J Pharmacol Sci. 2007 Jan;103(1):103-12.
We established an experimental system for measuring blood flow in the rat fundus and examined whether intravenously administered vasodilatory prostaglandins (PGE(1), PGE(2), and PGI(2)), 8-(4-chlorophenylthio)-cAMP (a cAMP analogue), and nicardipine (a Ca(2+)-channel blocker) increase fundus blood flow (FBF). Under artificial ventilation, rats were injected with tetrodotoxin (50 microg/kg, i.v.) to eliminate any nerve activity and prevent movement of the eye. After tetrodotoxin, the rats were infused with norepinephrine (0.3 - 0.5 microg . kg(-1) . min(-1)) and epinephrine (2.7 - 4.5 microg . kg(-1) . min(-1)) simultaneously to maintain adequate systemic circulation. We found that intravenous infusion of PGE(1) (2 - 10 microg . kg(-1) . min(-1)), PGE(2) (3 - 30 microg . kg(-1) . min(-1)), and PGI(2) (1 - 10 microg . kg(-1) . min(-1)) increased the FBF in a dose-dependent manner. The vasodilatory PGs decreased arterial pressure, whereas they did not affect heart rate. Like vasodilatory PGs, 8-(4-chlorophenylthio)-cAMP (30 micromol/kg, i.v.) increased FBF and decreased arterial pressure. While infusion of nicardipine (0.3 - 3 microg . kg(-1) . min(-1)) produced comparable depressor responses with those to vasodilatory PGs and the cAMP analogue, it did not increase FBF. These results suggest that vasodilatory PGs and cAMP act more selectively than Ca(2+)-channel blockers on retinal/choroidal blood vessels. Therefore, the vasodilatory PGs might be considered to be possible candidates for the therapeutics to treat disorders of retinal/choroidal circulation. [Abstract/Link to Full Text]

Ihara A, Wada K, Yoneda M, Fujisawa N, Takahashi H, Nakajima A
Blockade of leukotriene B4 signaling pathway induces apoptosis and suppresses cell proliferation in colon cancer.
J Pharmacol Sci. 2007 Jan;103(1):24-32.
We investigated whether leukotriene B(4) (LTB(4)) and its signaling pathway play an important role in the progression of human colon cancer via a direct stimulation of cancer cell proliferation. Remarkable expression of LTB(4) receptor 1 (BLT1) in human colon cancer tissues was detected by immunohistochemistry, and Western blot analysis revealed the BLT1 expression in cultured human colon cancer cell lines, Caco2 and HT29. The 5-lipoxygenase inhibitor AA-861 and LTB(4)-receptor antagonist U75302 showed negative effects on survival and proliferation of both Caco2 and HT-29 cells. The inhibition of cell proliferation is due to the apoptosis because nuclear condensation and increased annexin V expression were observed in the cells treated with AA-861 and U75302. Knockdown of BLT1 by small interfering RNA caused the suppression of BLT1 protein, resulting in the inhibition of cancer cell proliferation. Blockade of BLT1 by the receptor antagonist significantly suppresses the LTB(4)-stimulated extracellular signal-regulated kinase (ERK) activation in colon cancer cells. These results indicate that the blockade of the LTB(4)-signaling pathway induces apoptosis via the inhibition of ERK activation in colon cancer cells. The LTB(4)-signaling pathway might be a new therapeutic target for colon cancer. [Abstract/Link to Full Text]

Recent Articles in Alternative Medicine Review

Kidd P
Th1/Th2 balance: the hypothesis, its limitations, and implications for health and disease.
Altern Med Rev. 2003 Aug;8(3):223-46.
One theory of immune regulation involves homeostasis between T-helper 1 (Th1) and T-helper 2 (Th2) activity. The Th1/Th2 hypothesis arose from 1986 research suggesting mouse T-helper cells expressed differing cytokine patterns. This hypothesis was adapted to human immunity, with Th1- and Th2-helper cells directing different immune response pathways. Th1 cells drive the type-1 pathway ("cellular immunity") to fight viruses and other intracellular pathogens, eliminate cancerous cells, and stimulate delayed-type hypersensitivity (DTH) skin reactions. Th2 cells drive the type-2 pathway ("humoral immunity") and up-regulate antibody production to fight extracellular organisms; type 2 dominance is credited with tolerance of xenografts and of the fetus during pregnancy. Overactivation of either pattern can cause disease, and either pathway can down-regulate the other. But the hypothesis has major inconsistencies; human cytokine activities rarely fall into exclusive pro-Th1 or -Th2 patterns. The non-helper regulatory T cells, or the antigen-presenting cells (APC), likely influence immunity in a manner comparable to Th1 and Th2 cells. Many diseases previously classified as Th1 or Th2 dominant fail to meet the set criteria. Experimentally, Th1 polarization is readily transformed to Th2 dominance through depletion of intracellular glutathione, and vice versa. Mercury depletes glutathione and polarizes toward Th2 dominance. Several nutrients and hormones measurably influence Th1/Th2 balance, including plant sterols/sterolins, melatonin, probiotics, progesterone, and the minerals selenium and zinc. The long-chain omega-3 fatty acids EPA (eicosapentaenoic acid) and DHA (docosahexaenoic acid) significantly benefit diverse inflammatory and autoimmune conditions without any specific Th1/Th2 effect. Th1/Th2-based immunotherapies, e.g., T-cell receptor (TCR) peptides and interleukin-4 (IL-4) injections, have produced mixed results to date. [Abstract/Link to Full Text]

Czap A
Pass the tablet please.
Altern Med Rev. 2003 Aug;8(3):222. [Abstract/Link to Full Text]

Betaine. Monograph.
Altern Med Rev. 2003 May;8(2):193-6. [Abstract/Link to Full Text]

Humulus lupus. Monograph.
Altern Med Rev. 2003 May;8(2):190-2. [Abstract/Link to Full Text]

Cimicifuga racemosa. Monograph.
Altern Med Rev. 2003 May;8(2):186-9. [Abstract/Link to Full Text]

Prousky JE
Pellagra may be a rare secondary complication of anorexia nervosa: a systematic review of the literature.
Altern Med Rev. 2003 May;8(2):180-5.
Pellagra is a nutritional wasting disease attributable to a combined deficiency of tryptophan and niacin (nicotinic acid). It is characterized clinically by four classic symptoms often referred to as the four Ds: diarrhea, dermatitis, dementia, and death. Prior to the development of these symptoms, other nonspecific symptoms insidiously manifest and mostly affect the dermatological, neuropsychiatric, and gastrointestinal systems. A review of the literature reveals several case reports describing pellagra in patients with anorexia nervosa. The most common features of pellagra in patients with anorexia nervosa are cutaneous manifestations such as erythema on sun-exposed areas, glossitis, and stomatitis. Health care providers might consider a trial of 150-500 mg niacin if anorexic patients exhibit these cutaneous findings. Pellagra can be diagnosed if cutaneous symptoms resolve within 24-48 hours after oral niacin administration. To further corroborate a diagnosis of pellagra in anorexic patients, specific 24-hour urine tests for niacin metabolites and 5-hydroxy-indole-acetic acid could be run prior to treatment with niacin being instituted. Other factors, such as mycotoxins, excessive dietary leucine intake (although not in anorexia), estrogens and progestogens, carcinoid syndrome, and various medications, might also lead to the development of pellagra. Although pellagra appears to be a rare, yet possible secondary complication of anorexia nervosa, it should be considered in the work-up of patients who exhibit cutaneous manifestations subsequent to sunlight exposure. [Abstract/Link to Full Text]

Sampalis F, Bunea R, Pelland MF, Kowalski O, Duguet N, Dupuis S
Evaluation of the effects of Neptune Krill Oil on the management of premenstrual syndrome and dysmenorrhea.
Altern Med Rev. 2003 May;8(2):171-9.
PRIMARY OBJECTIVE: To evaluate the effectiveness of Neptune Krill Oil (NKO) for the management of premenstrual syndrome and dysmenorrhea. SECONDARY OBJECTIVE: To compare the effectiveness of NKO for the management of premenstrual syndrome and dysmenorrhea with that of omega-3 fish oil. METHODS/ DESIGN: Double-blind, randomized clinical trial. SETTING: Outpatient clinic. PARTICIPANTS: Seventy patients of reproductive age diagnosed with premenstrual syndrome according to the Diagnostic and Statistical Manual of Mental Disorders, Third Edition, Revised (DSM-III-R). INTERVENTIONS: Treatment period of three months with either NKO or omega-3 fish oil. OUTCOME MEASURES: Self-Assessment Questionnaire based on the American College of Obstetricians and Gynecologists (ACOG) diagnostic criteria for premenstrual syndrome and dysmenorrhea and number of analgesics used for dysmenorrhea. RESULTS: In 70 patients with complete data, a statistically significant improvement was demonstrated among baseline, interim, and final evaluations in the self assessment questionnaire (P < 0.001) within the NKO group as well as between-group comparison to fish oil, after three cycles or 45 and 90 days of treatment. Data analysis showed a significant reduction of the number of analgesics used for dysmenorrhea within the NKO group (comparing baseline vs. 45- vs. 90-day visit). The between-groups analysis illustrated that women taking NKO consumed significantly fewer analgesics during the 10-day treatment period than women receiving omega-3 fish oil (P < 0.03). CONCLUSION: Neptune Krill Oil can significantly reduce dysmenorrhea and the emotional symptoms of premenstrual syndrome and is shown to be significantly more effective for the complete management of premenstrual symptoms compared to omega-3 fish oil. [Abstract/Link to Full Text]

Marshall K
Cervical dysplasia: early intervention.
Altern Med Rev. 2003 May;8(2):156-70.
Cervical cancer is the second-most common cancer in young women and is one of the most common causes of cancer deaths among women, particularly in minorities and in impoverished countries. Cervical dysplasia, a premalignant lesion that can progress to cervical cancer, is caused primarily by a sexually transmitted infection with an oncogenic strain of the human papillomavirus (HPV). Not all women with the virus develop cervical dysplasia or cervical cancer. It has been postulated there are multiple host factors that contribute to progression of disease. Many of these factors, such as nutrient deficiencies, can be reversed, which will result in regression of dysplastic lesions. Studies have shown dietary intervention and nutrient supplementation to be effective in preventing cervical cancer. Additionally, local escharotic treatment combined with systemic treatment shows significant potential in reducing dysplasia. Recent advances in vaccination technology demonstrate the effectiveness of an HPV vaccine. The vaccine, however, may have many social and cost-prohibiting limitations, as well as health side effects. [Abstract/Link to Full Text]

Drisko JA, Giles CK, Bischoff BJ
Probiotics in health maintenance and disease prevention.
Altern Med Rev. 2003 May;8(2):143-55.
Probiotic microflora display numerous health benefits beyond providing basic nutritional value. They cooperatively maintain a delicate balance between the gastrointestinal tract and immune system. When this balance is disrupted, disease and inflammation result. Inflammation and over stimulation of the immune system by pathogenic bacteria are competitively inhibited by mucosal adherence of normal beneficial microflora. A healthy gastrointestinal tract with adequate mucus production and appropriate bacterial colonization prevents the overgrowth of pathogenic bacteria, modulates disease processes, and prevents widespread inflammatory disorders. The understanding of the function of probiotics in the maintenance of health and their importance in preventing disease serves to enhance the overall health of patients. With increasing understanding that beneficial microbes are required for health maintenance and disease prevention, probiotics may be commonly used as a therapeutic tool by health care practitioners in the not-too-distant future. This review presents a review of probiotics in health maintenance and disease prevention. [Abstract/Link to Full Text]

Hawrelak J
Giardiasis: pathophysiology and management.
Altern Med Rev. 2003 May;8(2):129-42.
Giardia, a common human parasite, can cause significant morbidity; however, natural medicine has great potential to influence the course of Giardia infection. The most beneficial way to treat giardiasis naturally may be through a combination approach, utilizing both nutritional interventions and phytotherapeutic agents. Nutritional intervention aims to reduce the acute symptoms of Giardia and help clear the infection. This can best be achieved by consuming a whole-food based, high-fiber, diet that is low in fat, lactose, and refined sugars. Additionally, ingestion of probiotics and wheat germ assists in parasite clearance. Numerous medicinal herbs show promise in the treatment of giardiasis. Berberine-containing herbs, garlic, and the Ayurvedic formulation Pippali rasayana currently have the most clinical evidence supporting their use. Blending the nutritional interventions and phytotherapeutic agents outlined in this article can minimize Giardia symptomatology and aid clearance of the parasite, without significant ill effects. As such, this therapeutic strategy should be considered the first-line approach. Antibiotic use may best be reserved for cases that fail to respond to initial treatment with natural measures. [Abstract/Link to Full Text]

Patrick L
Toxic metals and antioxidants: Part II. The role of antioxidants in arsenic and cadmium toxicity.
Altern Med Rev. 2003 May;8(2):106-28.
Exposure to toxic metals has become an increasingly recognized source of illness worldwide. Both cadmium and arsenic are ubiquitous in the environment, and exposure through food and water as well as occupational sources can contribute to a well-defined spectrum of disease. The symptom picture of arsenic toxicity is characterized by dermal lesions, anemia, and an increased risk for cardiovascular disease, diabetes, and liver damage. Cadmium has a significant effect on renal function, and as a result alters bone metabolism, leading to osteoporosis and osteomalacia. Cadmium-induced genotoxicity also increases risk for several cancers. The mechanisms of arsenic- and cadmium-induced damage include the production of free radicals that alter mitochondrial activity and genetic information. The metabolism and excretion of these heavy metals depend on the presence of antioxidants and thiols that aid arsenic methylation and both arsenic and cadmium metallothionein-binding. S-adenosylmethionine, lipoic acid, glutathione, selenium, zinc, N-acetylcysteine (NAC), methionine, cysteine, alpha-tocopherol, and ascorbic acid have specific roles in the mitigation of heavy metal toxicity. Several antioxidants including NAC, zinc, methionine, and cysteine, when used in conjunction with standard chelating agents, can improve the mobilization and excretion of arsenic and cadmium. [Abstract/Link to Full Text]

Czap A
Robert C. Atkins, MD, 1930-2003. In memoriam.
Altern Med Rev. 2003 May;8(2):105. [Abstract/Link to Full Text]

Astragalus membranaceus. Monograph.
Altern Med Rev. 2003 Feb;8(1):72-7. [Abstract/Link to Full Text]

Selenium. Monograph.
Altern Med Rev. 2003 Feb;8(1):63-71. [Abstract/Link to Full Text]

Thiamine. Monograph.
Altern Med Rev. 2003 Feb;8(1):59-62. [Abstract/Link to Full Text]

Lamson DW, Wright JV
A case of early renal functional impairment resolved with nutrients and botanicals.
Altern Med Rev. 2003 Feb;8(1):55-8.
The use of three herbal/nutritional products over a period of two months normalized blood urea nitrogen (BUN), serum creatinine, and creatinine clearance in a case of early functional kidney impairment. Although previous use of intravenous EDTA resolved Raynaud's syndrome symptoms, it provided little improvement to abnormal creatinine clearance. [Abstract/Link to Full Text]

Kelly G
The interaction of cigarette smoking and antioxidants. Part III: ascorbic acid.
Altern Med Rev. 2003 Feb;8(1):43-54.
The requirement for antioxidant nutrients depends on a person's exposure to endogenous and exogenous reactive oxygen species. Since cigarette smoking results in an increased cumulative exposure to reactive oxygen species from both sources, it would seem cigarette smokers might have an increased requirement for antioxidant nutrients. This review examines available evidence of ascorbic acid supplementation and combinations of antioxidants as interventions in smokers and their effect on functional biomarkers of nicotine metabolism, oxidative stress, DNA damage, and endothelial function. [Abstract/Link to Full Text]

MacKay D
Can CAM therapies help reduce antibiotic resistance?
Altern Med Rev. 2003 Feb;8(1):28-42.
The Centers for Disease Control and Prevention (CDC) reported the consumption of 235 million doses of antibiotics in 2001. It is estimated that 20-50 percent of these were unnecessarily prescribed for viral infections. Bacteria that antibiotics have controlled in the past are increasingly developing resistance to these drugs. Today, virtually all important bacterial infections in the United States and throughout the world are becoming resistant. For this reason, antibiotic resistance is among the CDC's top concerns. A large portion of antibiotics are dispensed by pediatricians treating common outpatient infectious diseases. The overuse of antimicrobials is beginning to be discouraged as scientific evidence is emerging to support the use of other therapies. In pediatric practice an emphasis on accurate diagnoses, control of environmental risk factors, and utilization of complementary and alternative medicine (CAM) therapies could reduce antibiotic prescribing. Antibiotic resistance poses a growing threat to health. CAM therapies may provide a safer, more effective treatment for many acute infections of childhood. [Abstract/Link to Full Text]

Basch E, Ulbricht C, Kuo G, Szapary P, Smith M
Therapeutic applications of fenugreek.
Altern Med Rev. 2003 Feb;8(1):20-7.
Fenugreek has a long history of medical uses in Ayurvedic and Chinese medicine, and has been used for numerous indications, including labor induction, aiding digestion, and as a general tonic to improve metabolism and health. Preliminary animal and human trials suggest possible hypoglycemic and antihyperlipidemic properties of oral fenugreek seed powder. [Abstract/Link to Full Text]

Miller AL
The methionine-homocysteine cycle and its effects on cognitive diseases.
Altern Med Rev. 2003 Feb;8(1):7-19.
Homocysteine, a sulfur-containing amino acid, is a metabolite of the essential amino acid methionine, and exists at a critical biochemical intersection in the methionine cycle - between S-adenosylmethionine, the indispensable ubiquitous methyl donor, and vitamins B12 and folic acid. High blood levels of homocysteine signal a breakdown in this vital process, resulting in far-reaching biochemical and life consequences. The link between homocysteine and cardiovascular disease is well established, and decreasing plasma total homocysteine by providing nutritional cofactors for its metabolism has been shown to reduce the risk of cardiovascular events. Information has been emerging regarding a connection between homocysteine metabolism and cognitive function, from mild cognitive decline (age-related memory loss) to vascular dementia and Alzheimer's disease. Significant deficiencies in the homocysteine re-methylation cofactors cobalamin (B12) and folate, as well as the trans-sulfuration cofactor vitamin B6, are commonly seen in the elderly population, with a resultant increase in homocysteine with advancing age. Hyperhomocysteinemia has been shown to be an independent risk factor for cognitive dysfunction. Indirect and direct vascular damage can be caused by homocysteine, which has been implicated in vascular dementia, with an increased risk of multiple brain infarcts and dementia as homocysteine levels rise. A significant correlation has been found between risk of Alzheimer's disease and high plasma levels of homocysteine, as well as low levels of folic acid, and vitamins B6 and B12. All of these disease associations are thought to be interrelated via increased homocysteine and S-adenosylhomocysteine and subsequent hypomethylation of numerous substances, including DNA and proteins, that render vascular structures and neurons more susceptible to damage and apoptosis. Providing the nutritional cofactors for proper functioning of the methionine cycle may improve methylation and protect the brain from damage. Further studies need to be performed to assess whether this will also reduce the risk of cognitive diseases and/or improve cognitive functioning. [Abstract/Link to Full Text]

Niacinamide. Monograph.
Altern Med Rev. 2002 Dec;7(6):525-9. [Abstract/Link to Full Text]

Isatis tinctoria. Monograph.
Altern Med Rev. 2002 Dec;7(6):523-4. [Abstract/Link to Full Text]

Appleton J
Arginine: Clinical potential of a semi-essential amino.
Altern Med Rev. 2002 Dec;7(6):512-22.
Arginine, a semi-essential amino acid, is involved in numerous areas of human biochemistry, including ammonia detoxification, hormone secretion, and immune modulation. Arginine is also well known as a precursor to nitric oxide (NO), a key component of endothelial-derived relaxing factor, an endogenous messenger molecule involved in a variety of endothelium-dependent physiological effects in the cardiovascular system. Because of arginine's NO-stimulating effects, it can be utilized in therapeutic regimens for angina pectoris, congestive heart failure, hypertension, coronary heart disease, preeclampsia, intermittent claudication, and erectile dysfunction. In addition, arginine has been studied in the treatment of HIV/AIDS, athletic performance, burns and trauma, cancer, diabetes and syndrome X, gastrointestinal diseases, male and female infertility, interstitial cystitis, immunomodulation, and senile dementia. Toxicity, dosage considerations, and contraindications are also reviewed. [Abstract/Link to Full Text]

Kelly GS
The interaction of cigarette smoking and antioxidants. Part 2: alpha-tocopherol.
Altern Med Rev. 2002 Dec;7(6):500-11.
It is logical that the requirement for antioxidant nutrients depends on a person's exposure to endogenous and exogenous reactive oxygen species. Since cigarette smoking results in an increased cumulative exposure to reactive oxygen species, it would seem cigarette smokers would have an increased requirement for antioxidant nutrients. This review examines available evidence of alpha-tocopherol supplementation by smokers and its effect clinically and on in vitro biomarkers of oxidative stress. [Abstract/Link to Full Text]

Kidd PM
Autism, an extreme challenge to integrative medicine. Part 2: medical management.
Altern Med Rev. 2002 Dec;7(6):472-99.
Autism and allied autistic spectrum disorders (ASD) present myriad behavioral, clinical, and biochemical abnormalities. Parental participation, advanced testing protocols, and eclectic treatment strategies have driven progress toward cure. Behavioral modification and structured education are beneficial but insufficient. Dietary restrictions, including removal of milk and other casein dairy products, wheat and other gluten sources, sugar, chocolate, preservatives, and food coloring are beneficial and prerequisite to benefit from other interventions. Individualized IgG or IgE testing can identify other troublesome foods but not non-immune mediated food sensitivities. Gastrointestinal improvement rests on controlling Candida and other parasites, and using probiotic bacteria and nutrients to correct dysbiosis and decrease gut permeability. Detoxification of mercury and other heavy metals by DMSA/DMPS chelation can have marked benefit. Documented sulfoxidation-sulfation inadequacies call for sulfur-sulfhydryl repletion and other liver p450 support. Many nutrient supplements are beneficial and well tolerated, including dimethylglycine (DMG) and a combination of pyridoxine (vitamin B6) and magnesium, both of which benefit roughly half of ASD cases. Vitamins A, B3, C, and folic acid; the minerals calcium and zinc; cod liver oil; and digestive enzymes, all offer benefit. Secretin, a triggering factor for digestion, is presently under investigation. Immune therapies (pentoxifyllin, intravenous immunoglobulin, transfer factor, and colostrum) benefit selected cases. Long-chain omega-3 fatty acids offer great promise. Current pharmaceuticals fail to benefit the primary symptoms and can have marked adverse effects. Individualized, in-depth clinical and laboratory assessments and integrative parent-physician-scientist cooperation are the keys to successful ASD management. [Abstract/Link to Full Text]

Patrick L
Mercury toxicity and antioxidants: Part 1: role of glutathione and alpha-lipoic acid in the treatment of mercury toxicity.
Altern Med Rev. 2002 Dec;7(6):456-71.
Mercury exposure is the second-most common cause of toxic metal poisoning. Public health concern over mercury exposure, due to contamination of fish with methylmercury and the elemental mercury content of dental amalgams, has long been a topic of political and medical debate. Although the toxicology of mercury is complex, there is evidence for antioxidant protection in the prevention of neurological and renal damage caused by mercury toxicity. Alpha-lipoic acid, a coenzyme of pyruvate and alpha-ketoglutarate dehydrogenase, has been used in Germany as an antioxidant and approved treatment for diabetic polyneuropathy for 40 years. Research has attempted to identify the role of antioxidants, glutathione and alpha-lipoic acid specifically, in both mitigation of heavy metal toxicity and direct chelation of heavy metals. This review of the literature will assess the role of glutathione and alpha-lipoic acid in the treatment of mercury toxicity. [Abstract/Link to Full Text]

Rhodiola rosea. Monograph.
Altern Med Rev. 2002 Oct;7(5):421-3. [Abstract/Link to Full Text]

Medium chain triglycerides. Monograph.
Altern Med Rev. 2002 Oct;7(5):418-20. [Abstract/Link to Full Text]

Logan AC, Beaulne TM
The treatment of small intestinal bacterial overgrowth with enteric-coated peppermint oil: a case report.
Altern Med Rev. 2002 Oct;7(5):410-7.
Recent investigations have shown that bacterial overgrowth of the small intestine is associated with a number of functional somatic disorders, including irritable bowel syndrome (IBS), fibromyalgia, and chronic fatigue syndrome. A number of controlled studies have shown that enteric-coated peppermint oil (ECPO) is of benefit in the treatment of IBS. However, despite evidence of strong antimicrobial activity, ECPO has not been specifically investigated for an effect on small intestinal bacterial overgrowth (SIBO). A case report of a patient with SIBO who showed marked subjective improvement in IBS-like symptoms and significant reductions in hydrogen production after treatment with ECPO is presented. While further investigation is necessary, the results in this case suggest one of the mechanisms by which ECPO improves IBS symptoms is antimicrobial activity in the small intestine. [Abstract/Link to Full Text]

Gaby AR
Treatment with enteric-coated peppermint oil reduced small-intestinal bacterial overgrowth in a patient with irritable bowel syndrome.
Altern Med Rev. 2003 Feb;8(1):3; author reply 4-5. [Abstract/Link to Full Text]

Steriti R
Nutritional support for chronic myelogenous and other leukemias: a review of the scientific literature.
Altern Med Rev. 2002 Oct;7(5):404-9.
Chronic myelogenous leukemia (CML) is a slowly progressive disease characterized by the overproduction of granulocytes (neutrophils, eosinophils, and basophils). A blood smear shows moderate elevations in white blood cell counts that may persist for years and be benign. Platelets are increased in number, although their function is impaired, resulting in symptoms of easy bleeding (purpura, swollen gums). Conventional medical treatment is a marrow transplant and alkylating agents, which are usually prescribed only during crisis. Several nutrients and botanicals have been studied for use in CML, including vitamin A and all-trans retinoic acid (Retin-A), vitamin D3, vitamin E, vitamin B12, indirubin (found in herbs including Indigofera tinctoria and Isatis tinctoria), and Curcuma longa. This article briefly reviews the scientific literature on the therapeutic use of these nutrients for CML. [Abstract/Link to Full Text]

Gaby AR
Intravenous nutrient therapy: the "Myers' cocktail".
Altern Med Rev. 2002 Oct;7(5):389-403.
Building on the work of the late John Myers, MD, the author has used an intravenous vitamin-and-mineral formula for the treatment of a wide range of clinical conditions. The modified "Myers' cocktail," which consists of magnesium, calcium, B vitamins, and vitamin C, has been found to be effective against acute asthma attacks, migraines, fatigue (including chronic fatigue syndrome), fibromyalgia, acute muscle spasm, upper respiratory tract infections, chronic sinusitis, seasonal allergic rhinitis, cardiovascular disease, and other disorders. This paper presents a rationale for the therapeutic use of intravenous nutrients, reviews the relevant published clinical research, describes the author's clinical experiences, and discusses potential side effects and precautions. [Abstract/Link to Full Text]

Kelly GS
The interaction of cigarette smoking and antioxidants. Part I: diet and carotenoids.
Altern Med Rev. 2002 Oct;7(5):370-88.
It is logical that the requirement for antioxidant nutrients depends on a person's exposure to endogenous and exogenous reactive oxygen species. Since cigarette smoking results in an increased cumulative exposure to reactive oxygen species from both sources, it would seem cigarette smokers would have an increased requirement for antioxidant nutrients. Logic dictates that a diet high in antioxidant-rich foods such as fruits, vegetables, and spices would be both protective and a prudent preventive strategy for smokers. This review examines available evidence of fruit and vegetable intake, and supplementation of antioxidant compounds by smokers in an attempt to make more appropriate nutritional recommendations to this population. [Abstract/Link to Full Text]

Recent Articles in Journal of Physiology and Pharmacology

Nowak P, Szczerbak G, Biedka I, Drosik M, Kostrzewa RM, Brus R
Effect of ketanserin and amphetamine on nigrostriatal neurotransmission and reactive oxygen species in Parkinsonian rats. In vivo microdialysis study.
J Physiol Pharmacol. 2006 Dec;57(4):583-97.
5-HT(2A/2C) receptors are one of the most important in controlling basal ganglia outputs. In rodent models of Parkinson's disease (PD) blockade of these receptors increases locomotion and enhances the actions of dopamine (DA) replacement therapy. Moreover, previously we established that 5-HT(2A/2C) antagonist attenuate DA D(1) agonist mediated vacuous chewing movements (VCMs) which are considered as an animal representation of human dyskinesia. These findings implicate 5-HT neuronal phenotypes in basal ganglia pathology, and promote 5-HT(2) antagonists as a rational treatment approach for dyskinesia that is prominent in most instances of PD replacement therapy. In the current study we determined whether ketanserin (KET) and/or amphetamine (AMPH) affected dopaminergic neurotranssmision in intact and fully DA-denervated rats. Moreover, we looked into extraneuronal content of HO. of the neostriatum after AMPH and/or KET injection, assessed by HPLC analysis of dihydroxybenzoic acids (2,3- and 2, 5-DHBA) - spin trap products of salicylate. Findings from the present study demonstrated that there are no substantial differences in extraneuronal HO. generation in the neostriatum between control and parkinsonian rats. KET did not affect DA release in the fully DA-denervated rat's neostriatum and also did not enhance HO. production. As 5-HT(2A/2C) receptor-mediated transmission might prove usefulness not only in addressing motor complications of PD patients (dyskinesia) but also in addressing non-motor problems such depression and/or L-DOPA evoked psychosis, the findings from the current study showed that the use of 5-HT(2A/2C) receptor antagonists in Parkinson's disease does not impend the neostriatal neuropil to be damaged by these drugs. We concluded that 5-HT(2A/2C) receptor antagonists may provide an attractive non-dopaminergic target for improving therapies for some basal ganglia disorders. [Abstract/Link to Full Text]

Krzemi?ski K, Mikulski T, Nazar K
Effect of prolonged dynamic exercise on plasma adrenomedullin concentration in healthy young men.
J Physiol Pharmacol. 2006 Dec;57(4):571-81.
The aim of the study was to find out whether prolonged exercise influences plasma adrenomedullin (ADM) concentration and whether it is related to the hormonal, metabolic and cardiovascular changes. Eighteen healthy subjects (age 25+/-1 yrs) were submitted to cycle exercise for 90 min at 70% of maximal oxygen uptake. Heart rate (HR) and blood pressure (BP) were measured continously. Before, at 30(th) min, and at the end of exercise venous blood samples were taken for [ADM], noradrenaline [NA], adrenaline [A], atrial natriuretic peptide [ANP], plasma renin activity PRA, interleukin-6 [IL-6] and lactate [LA] determination. Significant increases in plasma ADM and IL-6 were found at 90(th) min whereas other hormones were elevated already at 30(th) min of exercise. Positive correlations were ascertained between [ADM] and [NA] (r=0.47), [ANP] (r=0.35) or [IL-6] (r=0.35) and between exercise-induced increases in [ADM] and [NA] (r=0.38). PRA correlated positively with [NA] and [ANP]. Negative correlation was found between plasma [ADM] and diastolic BP. The present data suggest that increase in sympathetic nervous activity and cytokine induction during prolonged exercise may be involved in plasma ADM release and that increase in ADM and ANP secretion may be a compensatory mechanism against further elevation of blood pressure. [Abstract/Link to Full Text]

Duda M, Czarnowska E, Kurzelewski M, Konior A, Beresewicz A
Ischemic preconditioning prevents endothelial dysfunction, P-selectin expression, and neutrophil adhesion by preventing endothelin and O2- generation in the post-ischemic guinea-pig heart.
J Physiol Pharmacol. 2006 Dec;57(4):553-69.
Evidence indicates that ischemia/reperfusion (IR) results in endothelial dysfunction and neutrophil adhesion in the post-ischemic myocardium and that ischemic preconditioning (IPC), superoxide dismutase (SOD), and anti-endothelin-1 (ET-1) interventions prevent these effects. We tested the hypothesis that ET-1-induced superoxide (O(2)(-)) generation mediates endothelial injury and neutrophil accumulation in the IR heart, that IPC protects the endothelium and prevents the adhesion by attenuating post-ischemic ET-1, and thus O(2)(-), generation, and that the mitochondrial ATP-dependent potassium channel (mK(ATP)) triggers the IPC-induced protection. Langendorff-perfused guinea-pig hearts were subjected either to 30 min ischemia/35 min reperfusion (IR) or were preconditioned prior to IR with three cycles of either 5 min ischemia/5 min reperfusion or 5 min infusion/5 min wash-out of mK(ATP) opener diazoxide (0.5 microM). Neutrophils were infused to the hearts at 15-25 min of the reperfusion. Coronary flow responses to acetylcholine (ACh) and nitroprusside (SNP) served as measures of endothelium-dependent and -independent vascular function, respectively. Myocardial outflow of ET-1 and O(2)(-), P-selectin expression, neutrophil adhesion and functional recoveries were followed during reperfusion. IR augmented ET-1 and O(2)(-) outflow, P-selectin expression, and neutrophil adhesion, and impaired ACh response. These effects were attenuated or prevented by IPC and diazoxide, and 5-hydroxydecanoate (a selective mK(ATP) blocker) abolished the effects of IPC and diazoxide. SOD (150 U/ml) and tezosentan (5 nM, a mixed ET-1-receptor antagonist) mimicked the effects of IPC, although they had no effect on the ET-1 generation. The preventive effect of IPC, SOD and tezosentan on P-selectin expression preceded their effect on neutrophil adhesion. These data suggest that in guinea-pig heart: (i) ET-1-induced O(2)(-) generation mediates the post-ischemic endothelial dysfunction, P-selectin expression and neutrophil adhesion; (ii) IPC and diazoxide afford protection by attenuating the ET-1, and thus O(2)(-) generation; (iii) the mK(ATP) opening triggers the IPC protection; (iv) endothelial injury promotes post-ischemic neutrophil adhesion, but not vice versa. [Abstract/Link to Full Text]

Besse S, Bulteau AL, Boucher F, Riou B, Swynghedauw B, de Leiris J
Antioxidant treatment prevents cardiac protein oxidation after ischemia-reperfusion and improves myocardial function and coronary perfusion in senescent hearts.
J Physiol Pharmacol. 2006 Dec;57(4):541-52.
Cardiovascular ageing is associated with an increase in cardiac susceptibility to ischaemia and reperfusion and production of reactive oxygen species has been suspected to be responsible for this age-associated particular vulnerability. To determine whether administration of antioxidant treatment could afford some protection against ischaemia and reperfusion during aging, isolated perfused hearts from adult and senescent rats were submitted to normoxia (180 min), prolonged low-flow ischaemia (15% of initial coronary flow;180 min) or low-flow ischaemia/reperfusion (45 min/30 min), without or with antioxidant enzymes (superoxide dismutase+catalase; 50IU/ml). Contractile function and coronary perfusion were measured and protein oxidation was quantitated in left ventricle after normoxia, ischaemia and ischaemia/reperfusion. Protein oxidation was higher in senescent than in adult hearts after ischaemia-reperfusion, in contrast to prolonged ischaemia. During prolonged ischaemia, antioxidant treatment prevented coronary vasoconstriction at both ages and delayed contractile dysfunction in senescent hearts but did not limit protein oxidation. During reperfusion, antioxidant treatment prevented coronary vasoconstriction and protein oxidation at both ages and considerably improved recovery of contractile function in senescent hearts. In conclusion, antioxidant treatment fully protects the senescent heart against ischaemia/reperfusion but not against prolonged ischaemia injury, indicating that oxidative stress plays a central role in the age-associated vulnerability to ischaemia-reperfusion. [Abstract/Link to Full Text]

Kramkowski K, Mogielnicki A, Buczko W
The physiological significance of the alternative pathways of angiotensin II production.
J Physiol Pharmacol. 2006 Dec;57(4):529-39.
Although the use of angiotensin converting enzyme inhibitors (ACE-Is) in clinical practice brought the great chance to recognize the RAS role in the physiology and pathology, there are still many questions which we cannot answer. This article reviews actually known pathways of angiotensin II (Ang II) and other peptides of renin-angiotensin system (RAS) production and their physiological significance. The various carboxy- and aminopeptidases generate a range of peptides, like Ang II, Ang III, Ang IV, Ang-(1-7) and Ang-(1-9) possessing their own and known biological activity. In this issue especially the alternative pathways of Ang II synthesis involving enzymes other than angiotensin-converting enzyme (ACE) are discussed. We present many evidences for the significance of a new pathway of Ang II production. It has been clearly shown that Ang I may be converted to Ang-(1-9) by angiotensin-converting enzyme-related carboxypeptidase (ACE-2) and then into Ang II in some tissues, but the enzymes responsible for this process are unknown till now. Although there are many data proving the existence of alternative pathways of Ang II production, we can still block only ACE and angiotensin receptor 1 (AT(1)) in clinical practice. It seems that a lot needs to be done before we can wildly complexively control RAS and treat more effectively cardiovascular disorders such as hypertension or heart failure. [Abstract/Link to Full Text]

Guzik TJ, Mangalat D, Korbut R
Adipocytokines - novel link between inflammation and vascular function?
J Physiol Pharmacol. 2006 Dec;57(4):505-28.
Obesity and obesity related diseases are a major public health problem. Recent studies have shown that fat tissue is not a simple energy storage organ, but exerts important endocrine and immune functions. These are achieved predominantly through release of adipocytokines, which include several novel and highly active molecules released abundantly by adipocytes like leptin, resistin, adiponectin or visfatin, as well as some more classical cytokines released possibly by inflammatory cells infiltrating fat, like TNF-alpha, IL-6, MCP-1 (CCL-2), IL-1. All of those molecules may act on immune cells leading to local and generalized inflammation and may also affect vascular (endothelial) function by modulating vascular nitric oxide and superoxide release and mediating obesity related vascular disorders (including hypertension, diabetes, atherosclerosis, and insulin resistance) but also cancer or non-alcoholic fatty liver diseases. Present review, in a concise form, focuses on the effects of major adipocytokines, characteristic for adipose tissue like leptin, adiponectin, resistin and visfatin on the immune system, particularly innate and adaptive immunity as well as on blood vessels. Macrophages and T cells are populating adipose tissue which develops into almost an organized immune organ. Activated T cells further migrate to blood vessels, kidney, brain and other organs surrounded by infiltrated fat leading to their damage, thus providing a link between metabolic syndrome, inflammation and cardiovascular and other associated disorders. Ceretain treatments may lead to significant changes in adipocytokine levels. For example include beta-2 adrenoreceptor agonists, thiazolidinediones as well as androgens lead to decrease of plasma leptin levels. Moreover future treatments of metabolic system associated disorders should focus on the regulation of adipocytokines and their modes of action. [Abstract/Link to Full Text]

Torli?ska K, Grochowalska A, Kupsz J, Skoracka J, Kojo S
In vivo and in vitro effects of hyperglycemia on Na+ -K+, Ca+2, Mg+2-dependent ATPases activity in brain synaptosomes of aging rats.
J Physiol Pharmacol. 2006 Nov;57 Suppl 7145-58.
Cerebral metabolism of glucose, one of the determinants of tissue ATP level, is crucial for the CNS function. The activity of P-type pumps: Na(+), K(+)-ATPase, Ca(+2)-ATPase and Mg(+2)-ATPase were examined in rat brain synaptosomes to determine if changes in the enzyme activity related to aging are potentially associated with alterations in glucose homeostasis. Male Wistar rats (newborn, 3- and 18-month-old) were sacrificed by decapitation and synaptic plasma membranes were isolated from brains. In vivo study demonstrated that 18-month-old rats were characterized by hyperglycemia, hyperinsulinemia and increased total antyoxidative status (TAS) level. These conditions had a different impact on activities of the ATPases tested in vivo: only the activity of Ca(+2)-ATPase decreased whereas that of Mg(+2)-ATPase increased significantly. In vitro experiments, prior incubation of isolated synaptosomes with glucose of concentrations corresponding to normoglycemia in vivo (4.5 - 6.5 mM), stimulated Ca(+2)-ATPase activity, whereas higher glucose concentrations (10.0 - 12.5 mM) inhibited significantly the enzyme activity. The most sensitive to hyperglycemia appeared Na(+), K(+)-ATPase in old rats synaptosomes with the progressive decline starting at 6.5 mM glucose. The activity of Mg(+2)-ATPase was not inhibited in vitro even at high glucose concentrations that may explain the increased in vivo, activity of this enzyme in old, hyperglycemic rats. [Abstract/Link to Full Text]

Bonior J, Jaworek J, Konturek SJ, Pawlik WW
Leptin is the modulator of HSP60 gene expression in AR42J cells.
J Physiol Pharmacol. 2006 Nov;57 Suppl 7135-43.
Leptin, circulating protein involved in the control of body weight and energy expenditure received attention as a modulator of immune response of the organism. Leptin receptors have been detected in the pancreas and experimental studies have shown that leptin protects the pancreas against the damage induced by caerulein overstimulation. Heat shock proteins (HSP) are endogenous proteins produced by various cells exposed to high temperature or to the noxious agents. HSP protect the cells against various environmental and endogenous stressors. The implication of HSP60 in the leptin-induced pancreatic protection has not been examined yet. The aim of this study was: to investigate the changes of HSP60 mRNA signal in the pancreatic AR42J cells subjected to caerulein and leptin. AR42J cells were incubated in standart medium at 37 degrees C for: 0, 1, 3, 5, 12 or 24 h, under basal conditions. Incubation time of 3 h was selected for the next experiments. AR42J cells were incubated in presence of caerulein (10(-11), 10(-9) or 10(-7) M), leptin (10(-8) or 10(-6) M), or combination of above. Gene expression for HSP60 was determined by RT-PCR. The mRNA signal for HSP60 has been observed in AR42J pancreatic cells under basal conditions. Incubation of AR42J cells in presence of leptin (10(-8) or 10(-6) M) resulted in the significant increase of gene expression for HSP60 in both groups of AR42J cells. Caerulein stimulation reduced mRNA signal for HSP60. The strongest mRNA signal for HSP60 has been observed after the exposition of AR42J cells to combination of leptin and caerulein. We conclude that: 1. Gene expression for HSP60 has been detected in pancreatic AR42J cells under basal conditions. 2. HSP60 gene expression was significantly increased in pancreatic AR42J cells stimulated by leptin whereas caerulein reduced this signal. 3. The strongest gene expression for HSP60 has been detected in the cells incubated with combination of caerulein and leptin. [Abstract/Link to Full Text]

Godlewski MM, Slazak P, Zabielski R, Piastowska A, Gralak MA
Quantitative study of soybean-induced changes in proliferation and programmed cell death in the intestinal mucosa of young rats.
J Physiol Pharmacol. 2006 Nov;57 Suppl 7125-33.
The use of soybean in human and animal nutrition is limited because of high content of bioactive compounds: enzyme inhibitors, polyphenols, goitrogens, phytates, saponins, sugars, and agglutinins. The damage of intestinal mucosa structure was previously observed in animals fed soybean supplemented diets. Hence, the objectives of the presented study were to compare intensity of epithelium remodeling processes in different intestinal segments, and to evaluate the influence of the 1% of soybean dietary supplementation on the processes in intestinal mucosa. The experiment was performed on 30 Wistar rats fed AIN-93 based diets. Animals were divided randomly into three groups: control (CTRL), with 1% of raw soybean (RS) and with 1% of soaked and boiled soybean (BS). The samples of: duodenum (DUO), proximal jejunum (PROX), mid-jejunum (MID), distal-jejunum (DIST) and ileum (ILE) were collected. The following processes in these samples were evaluated: mitosis (Ki-67), apoptosis (Cpp32), autophagy (MAP I LC3) and DNA damage (p53). Present data show that modification of soybean by soaking and subsequent boiling markedly influences the enterocyte turnover in the small intestine mucosa. Increased mitotic ratio in the intestine of rats fed with boiled soybean masks the negative effects of soybean on the small intestine structure. [Abstract/Link to Full Text]

Popow A, Nowak D, Malicka-B?aszkiewicz M
Actin cytoskeleton and beta-actin expression in correlation with higher invasiveness of selected hepatoma Morris 5123 cells.
J Physiol Pharmacol. 2006 Nov;57 Suppl 7111-23.
Our studies were focused on the isolation and characterization of highly motile fraction of cells from hepatoma Morris 5123 population. Cells that underwent several migration cycles through Matrigel - coated filters were successfully cultured. The invasion index was determined by means of Matrigel invasion assay. Statically significant increase in the value of invasion factor for selected cells variant in comparison to the parental population was observed. The considerable changes in the cell shape were followed by the reorganization of the actin cytoskeleton structure including a dense subcortical congestion in the distribution of beta -actin isoform. The visualization of this protein in tumor cells was performed by immunostaining and scanning fluorescent confocal microscopy. The results were confirmed by densitometry analysis of Western blots. In addition, the increased state of actin polymerization in the cytoplasmic fraction of selected cells was determined as measured by filamentous to monomeric (F:G) actin ratio. Concluding, the selected fraction of hepatoma Morris 5123 cells with higher invasion capacity was characterized by rounded shape, remarkable increase of beta -actin level, its submembrane concentration as well as with the increased state of actin polymerization with respect to parental cells population. [Abstract/Link to Full Text]

Sadkowski T, Jank M, Oprzadek J, Motyl T
Age-dependent changes in bovine skeletal muscle transcriptomic profile.
J Physiol Pharmacol. 2006 Nov;57 Suppl 795-110.
The postnatal growth of muscle tissue occurs by hypertrophy comprising satellite cells proliferation, differentiation and protein turnover. The highest rate of skeletal muscle gains and protein synthesis in bulls occurs in the period between 180 and 360 days of postnatal life. However, genes which are responsible for quantitative and qualitative changes in skeletal muscle during this period are not identified up to date. The aim of our study was to compare the changes in transcriptomic profile of skeletal muscle (m. semitendinosus) in 12 Polish Black and White bulls between 6 and 12 month of life. For experimental purposes we used bovine cDNA microarray (the NBFGC EST collection) which contains 18,263 unique genes, derived from many different tissue types and various physiologically important states within these tissues. Our results revealed 53 genes which expression changed in the same manner depending on age of all examined pairs of animals. Thirty two of these genes showed at least 2-fold difference in expression between two analyzed age points. Age-dependent up-regulation was the most pronounced in the case of following genes: similar to MAD2L1 binding protein, similar to thymocyte protein thy28 isoform 1, similar to type I inositol-1,4,5-triphosphate 5-phosphatase, similar to nucleoside diphosphate kinase 6, proline rich 14, similar to transcription factor E2-alpha and phospholipase C gamma 1. The highest age-dependent decrease of the transcript was observed in the case of: similar to ubiquitin carboxy-terminal hydrolase L1, similar to latent TGF-beta binding protein 3 precursor, phospho-mannomutase 2, CD74 antigen, similar to BCL6 co-repressor-like 1, platelet/endothelial cell adhesion molecule (PECAM1), necdin, zygin, tight junction protein 3, ankyrin and apolipoprotein-L3. Although the role of the most of above genes and interactions between products of their expression is not clear at the moment, the significance of their response between 6 and 12 month of age may indicate their involvement in growth, development and metabolic changes in bovine skeletal muscle during the first year of postnatal life. [Abstract/Link to Full Text]

Herosimczyk A, Dejeans N, Sayd T, Ozgo M, Skrzypczak WF, Mazur A
Plasma proteome analysis: 2D gels and chips.
J Physiol Pharmacol. 2006 Nov;57 Suppl 781-93.
The knowledge of concentration, modification and interaction of proteins is fundamental in determining the phenotype of living organisms. Plasma, the primary clinical specimen, contains numerous and diverse proteins. The functions of these proteins are as manifold as the diversity of the protein themselves. Many of them have been largely used for many years as biomarkers of diseases and indicators of the physiological functions. The study of plasma proteome promises to be a significant advance in various areas of biological and clinical research. Two-dimensional polyacrylamide gel electrophoresis is considered as a primary tool in separating thousand of plasma proteins. This approach enables comparing normal and diseased samples revealing differently expressed proteins. Other proteomic techniques suitable for plasma analysis such as protein microarrays are now either established or are still being improved. This article briefly reviews the application of two-dimensional electrophoresis and the current status of technical aspects for plasma proteome. [Abstract/Link to Full Text]

Kiela PR, Xu H, Ghishan FK
Apical NA+/H+ exchangers in the mammalian gastrointestinal tract.
J Physiol Pharmacol. 2006 Nov;57 Suppl 751-79.
The Slc9a family of nine Na(+)/H(+) exchangers (NHE) plays a critical role in neutral sodium absorption in the mammalian intestine as well as other absorptive and secretory epithelia of digestive organs. These transport proteins mediate the electroneutral exchange of Na(+) and H(+) and are crucial in a variety of physiological processes, including the fine tuning of intracellular pH, cell volume control and systemic electrolyte, acid-base and fluid volume homeostasis. Here, we review the role of the Na(+)/H(+) exchange mechanism as it relates to the physiology of organs and cells involved in nutrient absorption, and we describe physiological and molecular aspects of individual isoforms, including their structure, tissue-, and subcellular distribution, as well as their regulation by physiological stimuli at the transcriptional and post-transcriptional levels. A particular emphasis is placed on Na(+)/H(+) exchanger isoforms expressed on the apical (brush border) membrane of the epithelial cells, and the consequences of gene-targeted mutation of individual isoforms are discussed in the context of the physiology of digestive organs. Where available, we also provide a review of pathophysiological states related to aberrant expression and/or activity of Na(+)/H(+) exchangers within the confines of the digestive system. [Abstract/Link to Full Text]

Tudek B, Swoboda M, Kowalczyk P, Oli?ski R
Modulation of oxidative DNA damage repair by the diet, inflammation and neoplastic transformation.
J Physiol Pharmacol. 2006 Nov;57 Suppl 733-49.
Oxidative DNA damage and DNA repair may mediate several cellular processes, like replication and transcription, mutagenesis and apoptosis and thus may be important for the organism development as well as its pathogenesis, including cancer. Activity of DNA repair enzymes can depend on many factors, such as gene polymorphism, mRNA and protein level, as well as enzymes activation and inhibition. Modulation of base excision repair pathway eliminating from DNA oxidatively formed lesions may be caused by the diet, inflammation and neoplastic transformation. Reactive oxygen species and some diet components induce transcription of several Base Excision Repair enzymes, e.g. major human AP-endonuclease, (APE1) and 8-oxoG-DNA glycosylase (OGG1). The carcinogenic process in human lung decreases repair activity for 8-oxoGin transcription independent manner, but increases repair activity of epsilon A and epsilon C, as measured in tumors and unchanged lung tissues of lung cancer patients. Thus, modulation of repair enzymes activities may be a cell response on their way to differentiation ot neoplastic transformation. [Abstract/Link to Full Text]

Motyl T, Gajkowska B, Zarzy?ska J, Gajewska M, Lamparska-Przybysz M
Apoptosis and autophagy in mammary gland remodeling and breast cancer chemotherapy.
J Physiol Pharmacol. 2006 Nov;57 Suppl 717-32.
Apoptosis - programmed cell death (PCD) type I is physiological process responsible for cell loss during mammary gland involution after natural weaning or litter removal in rodents, after weaning in sow and during drying off in goat and cow. The regulation of mammary epithelial cell (MEC) apoptosis in bovine mammary gland occurs at three levels. The first level comprises intracellular regulatory proteins, e.g. Bcl-2 family death promoters and inhibitors. The second level is represented by intramammary inductors of apoptosis, e.g. FIL, IGFBPs, Fas ligand, TGF-betas. The expression and activity of these auto/paracrine inductors of apoptosis is controlled and modulated by the third level factors, e.g. systemic galactopoetic hormones, nutrition, reproductive status and milking management. Our recent study proved that apoptosis in involuting bovine mammary gland is accompanied by increased intensity of autophagy, regarded as a cytoprotective process but in advanced stage as a PCD type II. Moreover, we have reported for the first time the ability of TGF-beta(1) to induce both apoptosis and autophagy in bovine BME-UV1 MEC. Much more pronounced heterogeneity of PCD was observed when breast cancer cells were exposed to anticancer drugs. The primary responses of breast cancer MCF-7 cells to camptothecin (CPT) are apoptosis and autophagy (as a cytoprotective process). In this case autophagy occurs in cells which are resistant to apoptosis as a tool of cancer cell survival. The fail-safe responses of breast cancer cells to persisting CPT-induced stress are apoptosis accompanied by morphological and biochemical features of autophagy or type II PCD with advanced subcellular degradation. The threshold between autophagy as a cytoprotective process (reversible) or PCD (irreversible) is difficult to establish and probably depends on the extent of degradation of cellular components. Proapoptotic protein Bid may serve as a molecular switch between apoptosis and autophagy. Bid knock down in MCF-7 cells exposed to CPT leads to a shift of cell death from apoptosis to autophagy. Since bid and other proapoptotic genes undergo mutations in malignant cells, the ability of cancer cells commitment to autophagy may have important therapeutic implications. [Abstract/Link to Full Text]

Kucia M, Ratajczak MZ
Stem cells as a two edged sword--from regeneration to tumor formation.
J Physiol Pharmacol. 2006 Nov;57 Suppl 75-16.
Evidence has accumulated that quiescent stem cells or cells developmentally closely related to them distributed in various organs may be a cellular origin of cancer development. In support of this notion, stem cells (SC) are long-lived cells with distinctive properties of self-renewal and has the potential to proliferate extensively. Given these features, it's possible that they may become the subject of consecutive accumulated mutations that are crucial for initiation of cancer. Therefore, mutations that occur in normal stem cells might lead to their malignant transformation and tumor initiation. Furthermore, many biological features of normal and cancer SC such as the physiological trafficking of normal and metastasis of cancer stem cells involve similar molecular mechanisms, and we discuss these similarities here. [Abstract/Link to Full Text]

Kochan Z, Karbowska J, Swierczynski J
The effects of weight cycling on serum leptin levels and lipogenic enzyme activities in adipose tissue.
J Physiol Pharmacol. 2006 Nov;57 Suppl 6115-27.
Weight cycling is one of the widely used weight reduction strategies; however, the adverse effects of this method include regaining significant amounts of weight. The molecular mechanisms underlying weight gain following cycles of dietary deprivation and refeeding are still poorly understood. One of the possibilities is that repeated loss and gain of weight may promote fat deposition in adipose tissue. To test this hypothesis we investigated serum leptin levels and lipogenic enzyme activities in white adipose tissue (WAT) of male Wistar rats during 12 days of ad libitum feeding following multiple cycles of alternating food deprivation and refeeding. Rats subjected to eight cycles of food deprivation and refeeding (MFR group) showed significantly decreased circulating leptin levels when compared with control rats (nearly 50% decrease in leptin levels, P < 0.01). Throughout 12 days of ad libitum feeding, serum leptin levels increased modestly but remained significantly (24%, P < 0.05) lower than control levels. Fatty acid synthase (FAS) and malic enzyme (ME) activities (chosen as representatives of enzymes directly involved in fatty acid synthesis) were found to be considerably higher in WAT of MFR rats refed for 3 days in comparison to control rats, and remained elevated even after 12 days of refeeding. These observations suggest that the elevation of lipogenic enzyme activities induced by multiple cycles of dietary deprivation followed by refeeding persists for several days, markedly increasing the lipogenic capacity of adipose tissue, which, accompanied by a decrease in circulating leptin levels, may promote weight gain. [Abstract/Link to Full Text]

Karbowska J, Kochan Z
Role of adiponectin in the regulation of carbohydrate and lipid metabolism.
J Physiol Pharmacol. 2006 Nov;57 Suppl 6103-13.
Adiponectin, an adipocyte-derived plasma protein, has been shown to play an important role in the regulation of fatty acid and glucose metabolism. Adiponectin enhances fatty acid oxidation both in skeletal and cardiac muscle as well as in the liver, thus reducing triglyceride content in these tissues. Moreover, it stimulates glucose uptake by skeletal and cardiac muscle, and inhibits glucose production by the liver; consequently decreasing blood glucose levels. This review focuses on the molecular mechanisms underlying adiponectin effects on carbohydrate and lipid metabolism in skeletal muscle, cardiac muscle and liver. [Abstract/Link to Full Text]

Swierczynski J
Leptin and age-related down-regulation of lipogenic enzymes genes expression in rat white adipose tissue.
J Physiol Pharmacol. 2006 Nov;57 Suppl 685-102.
The age-related inverse relationship between gene expression of lipogenic enzymes and leptin gene expression as well as inhibitory effect of leptin on lipogenic enzyme's gene expression suggest that leptin could be responsible in part for the low rate of lipogenesis in white adipose (WAT) of old rats. Based on the data published recently we propose a model for the direct inhibitory effect of leptin on lipogenesis. This model may explain the age-related decrease of lipogenic activity in WAT. It is likely that despite of higher concentration of noradrenaline (which inhibits leptin gene expression in WAT) in old animals, the age-dependent decrease of beta-adrenergic receptor density in rat adipocytes may lead to the increase of leptin gene expression and to the increase of WAT leptin concentration. High concentration of leptin in adipose tissue decreases sterol regulatory element binding protein-1 (SREBP-1) gene expression by paracrine and/or autocrine action on adipocyte, which leads to the decrease of SREBP-1c level (mature form). The suppression of SREBP-1c synthesis causes a decrease of lipogenic enzyme's gene expression which consequently results in lower rate of fatty acid synthesis in WAT. This model does not exclude the indirect, via hypothalamus (by decreasing food consumption), inhibitory action of leptin on WAT lipogenesis. Therefore, it is likely that leptin exerts its inhibitory effect on WAT lipogenesis both directly at the level of adipocytes, and indirectly through hypothalamus by decreasing food intake. The inhibitory effect of of high leptin concentration on lipogenesis in WAT of old rats could prevent over-accumulation of triacylglycerols in adipocyte, and by this way could protect against further development of the fat mass. [Abstract/Link to Full Text]

Kmiec Z, Pokrywka L, Kotlarz G, Mysliwski A
The effects of fasting and refeeding on adrenal cortex morphometry and serum concentrations of ACTH and corticosterone in young and old male rats.
J Physiol Pharmacol. 2006 Nov;57 Suppl 677-84.
The impairment of homeostatic mechanisms in ageing becomes often apparent upon physiological or pathological stimulation. We have previously shown that fasting and refeeding revealed the existence of age-related changes of carbohydrate and lipid metabolism. Because fuel metabolism is partially controlled by corticosteroids we decided to determine the effects of refeeding on adrenal gland morphometry, ACTH, and corticosterone serum levels in young (5 mo) and (20 mo) old male Wistar rats. Fasting for 48 h did not change serum ACTH and corticosterone in both age groups. ACTH level did not change after 24 h of refeeding in young and old rats. However, in old, but not young animals, refeeding resulted in the decrease of corticosterone serum concentration. The relative weight of adrenal gland (% of body weight) did not change significantly with age (p=0.05). Fasting for 48 h induced in old rats but not in young ones increase of relative adrenal weight, and the volume of the reticular zone. Refeeding reduced adrenal volume, fascicular zone and reticular zone. Refeeding for 24 h decreased the total volume of adrenal gland of old rats due to a decline of the volumes of fascicular and reticular zones. In young rats refeeding reduced the volume of reticular zone. It is concluded that refeeding revealed ageing-dependent decline in the secretion of corticosterone, the key hormone of prolonged stress response. [Abstract/Link to Full Text]

Brzozowski T, Konturek PC, Sliwowski Z, Drozdowicz D, Kwiecien S, Pawlik M, Pajdo R, Konturek SJ, Pawlik WW, Hahn EG
Neural aspects of ghrelin-induced gastroprotection against mucosal injury induced by noxious agents.
J Physiol Pharmacol. 2006 Nov;57 Suppl 663-76.
Ghrelin, identified in oxyntic mucosa has been recently implicated in the control of food intake and growth hormone (GH) release but whether this hormone can influence the gastric secretion and gastric mucosal integrity have been little studied. We compared the effects of intraperitoneal (i.p.) and intracerebroventricular (i.c.v.) administration of ghrelin on gastric secretion in rats equipped with gastric fistula (GF) and gastric lesions induced in rats by 75% ethanol and ischemia-reperfusion (I/R) with or without vagotomy or functional ablation of afferent sensory nerves by capsaicin. The number and the area of gastric lesions was measured by planimetry, the GBF was assessed by H(2)-gas clearance method and blood was withdrawn for the determination of the plasma ghrelin and gastrin levels. Ghrelin (5-80 microg/kg i.p. or 600-5000 ng/rat i.c.v.) increased gastric acid secretion and attenuated gastric lesions induced by ethanol and I/R. These protective effects of ghrelin were accompanied by the significant rise in the gastric mucosal blood flow (GBF) and plasma ghrelin and gastrin levels. Ghrelin given i.p. or injected i.c.v. in standard doses 20 microg/kg or 5000 ng/kg, respectively, significantly attenuated the gastric mucosal damage and significantly raised the GBF. Ethanol applied i.g. in smaller concentrations (12.5% and 25%) produced a significant increase in plasma immunorective ghrelin levels and this effect was inhibited in rats receiving ethanol in higher concentrations (75% and 100%). Ghrelin-induced protection after its i.p. or i.c.v. administration and accompanying increase in the GBF were completely abolished by vagotomy and capsaicin-deactivation of sensory nerves. Concurrent treatment with CGRP added to ghrelin restored the gastroprotective and hyperemic effects of ghrelin applied i.p. or i.c.v. in rats with capsaicin denervation. We conclude that central and peripheral ghrelin exerts a potent protective and gastric secretory effects in rats exposed to ethanol and I/R, and that these actions involve vagal nerve integrity, partially depending upon afferent nerves and hyperemia mediated by sensory neuropeptides such as CGRP released from these nerves. [Abstract/Link to Full Text]

Baranowska B, Bik W, Baranowska-Bik A, Wolinska-Witort E, Szybinska A, Martynska L, Chmielowska M
Neuroendocrine control of metabolic homeostasis in Polish centenarians.
J Physiol Pharmacol. 2006 Nov;57 Suppl 655-61.
Neuropeptides play a pivotal role in the control of metabolic homeostasis. We aimed to evaluate the release of neuropeptides involved in the control of energy homeostasis in relation to metabolic status in aging humans. The study group consisted of 183 women: 75 centenarians (above 100 yrs old), 26 elderly women (below 70 yrs), 45 younger women (mean 26 yrs) and 37 obese women (mean 41.6 yrs). Fasting plasma concentration of leptin, adiponectin, ghrelin active, neuropeptide Y (NPY) and insulin were measured. Our results showed several differences in the metabolic and neurohormonal status in the centenarian group. The incidence of hypertension, glucose intolerance, insulin resistance and dyslipidemia was lower compared with obese women. Leptin and NPY concentrations were significantly lower than in elderly and obese subjects. Moreover, NPY level was higher than that in the younger group. Plasma adiponectin values were higher than in any of the other group. Insulin levels were significantly lower compared with the young and obese groups. Furthermore, a negative correlation was found between adiponectin and HOMA-IR, and adiponectin and insulin. Ghrelin active concentrations were significantly lower compared with the young subjects. However, ghrelin levels were higher than in obese subjects. We conclude that altered neuropeptide activity in centenarians may play a role in the mechanisms contributing to prolonged survival. [Abstract/Link to Full Text]

Korczynski W, Ceregrzyn M, Kato I, Wolinski J, Zabielski R
The effect of orexins on intestinal motility in vitro in fed and fasted rats.
J Physiol Pharmacol. 2006 Nov;57 Suppl 643-54.
Orexin-A and -B (OXA, OXB) are peptides involved in many gastrointestinal (GI) functions, including motility. Orexins, their precursors and receptors are present in the GI tract. The expression of orexins increases in the hypothalamus and gastrointestinal tract in response to fasting. We have examined the effect of OXA and OXB on GI motility in vitro in fed and fasted rats. The intestinal segments were mounted in chambers filled with Krebs solution. Isotonic contractions were measured in response to acetylcholine (10(-5) M), electric field stimulation (EFS), and orexins (10(-9)-10(-7) M) alone or in the presence of orexin-1 type receptor antagonist, SB- 334867 (10(-5) M), tetrodotoxin (TTX) 10(-6) M, or atropine (10(-5) M). Orexins caused a dose-dependent increase of intestinal segment contractions with a more pronounced effect of OXB over OXA. Fasting did not influence orexin-induced responses. Incubation with SB-334867 led to a marked decrease in orexin-induced contractions in OXA-treated segments, while those of OXB were not affected. Atropine diminished contractions only in fasted animals, while TTX led to a decreased response to orexins in both groups. The results show that OXB is predominant in inducing gut motility response, that the effect of orexins is not fully dependent on cholinergic and Na(+) transmissions, and that involvement of other transmitters is possible. [Abstract/Link to Full Text]

Korczynski W, Ceregrzyn M, Matyjek R, Kato I, Kuwahara A, Wolinski J, Zabielski R
Central and local (enteric) action of orexins.
J Physiol Pharmacol. 2006 Nov;57 Suppl 617-42.
Orexin-A (OXA, hypocretin-1) and orexin-B (OXB, hypocretin-2) are peptides derived from the same 130 amino acid long precursor (prepro-orexin) that bind and activate two closely related orphan G protein-coupled receptors. Orexins and their receptors were first discovered in the rat brain, and soon after that in peripheral neural structures, including the vagal nerve and enteric nervous system, and in other structures involving the gastrointestinal tract diffuse neuroendocrine system, pancreas tissue, stomach and intestinal mucosa. Orexins and their receptors were also demonstrated in the testes, adrenals, kidneys, and placenta. This review is focused on central and enteric actions. Originally, orexins were considered to be neurotransmitters that centrally stimulate food intake in animals and humans, but it soon became evident that their action is broader due to activation of a large number of neuronal pathways involved in energy homeostasis, sleep-awake behavior, nociception reward seeking, food and drug addiction, as well as reproduction, cardiovascular and adrenal function. In the gastrointestinal tract, orexins have been found so far to affect gastrointestinal motility and gastric, intestinal and pancreatic secretions. The effects were observed following central (intraventricular) or local (intraluminal, intraarterial), but not peripheral (intravenous), administrations of orexins. Since the expression of orexins in the gastrointestinal tract is enhanced during fasting, and fasting reveals many of the orexin gastrointestinal effects, it seems probable that on the local level, orexins keep the gastrointestinal tract functions ready during fasting and play role in brain-gut axis control. [Abstract/Link to Full Text]

Kmiec Z
Central regulation of food intake in ageing.
J Physiol Pharmacol. 2006 Nov;57 Suppl 67-16.
Energy homeostasis and fuel metabolism undergo significant modifications in the course of ageing. During the second half of life many humans increase their body mass and develop glucose intolerance that may lead to obesity and type 2 diabetes. However, many old people suffer from being underweight, and this "anorexia of elderly" may seriously compromise their health under certain circumstances. Experimental studies into the causes of ageing-related impairments of food intake regulation were performed mainly on rat, and to some extent, on non-human primates. It was found that the expression of NPY, the most potent orexigenic peptide, and of NPY receptors, is highly suppressed in the hypothalamus of old rats. Moreover, the increase of NPY mRNA after fasting was severely blunted in old as compared to young rats. Similar reductions, although of lower magnitude, were reported for other hypothalamic orexigenic compounds such as, AgRP and orexins. Interestingly, ageing does not significantly alter hypothalamic mRNA levels of important anorexigens such as CART and aMSH. The presented findings suggest that, at least in rodents, ageing is associated with the general down-regulation of hypothalamic peptides that stimulate food intake and unchanged expression of anorexigenic peptides. This situation may be responsible for the decreased appetite drive in senescent animals and loss of weight at the end-of-life period. If similar changes of the central control of food intake underly "anorexia of ageing" observed in some elderly, it is possible that therapeutic intervention at this regulatory level may be possible in the future. [Abstract/Link to Full Text]

Zayachkivska OS, Gzhegotsky MR, Terletska OI, Lutsyk DA, Yaschenko AM, Dzhura OR
Influence of Viburnum opulus proanthocyanidins on stress-induced gastrointestinal mucosal damage.
J Physiol Pharmacol. 2006 Nov;57 Suppl 5155-67.
Recent studies demonstrated that the proanthocyanidins (PA), the polymers of flavan-3-ols, naturally occurring plant metabolites widely available in fruits, vegetables, nuts, seeds, flowers and bark, have anti-inflammatory, anticarcinogenic, anti-allergic, antioxidant and vasodilatory actions. We hypothesized that Viburnum opulus PA (VOPA, Caprifoliaceae), due to activation of multifactorial gastrointestinal mucosal defense mechanisms, exert gastroduodenoprotective effects. The aim of the study was: 1) to investigate VOPA effects on gastroduodenal mucosal integrity and pattern of carbohydrate binding proteins and nitric oxide (NO) content in intact mucosa and that exposed to non-topical ulcerogens (stress) in rats without and with capsaicin (125 mg/kg, sc) denervation; and 2), to assess the role of activity of antioxidizing enzymes superoxide dismutase (SOD), catalase (CAT), gluthatione peroxidase (GPx) in VOPA-induced gastroduodenoprotection against water immersion and restraint stress (WRS) in rats. VOPA was administered orally in dose of 25, 50 or 75 mg/kg body weight. Gastroduodenal mucosal damage detected by routine light microscopic investigation and lectin histochemistry set, purified from plant and animal sources of Carpatian region. NO content, pro-and antioxidant system were determined by routine laboratory methods. Pretreatment with VOPA afforded gastroduodenoprotection and was accompanied by an increase in NO expression, both changes being reversed by sensory denervation, as well as by the rise of SOD, CAT activity and fall in MDA content. Our study shows that VOPA exerts a potent gastroduodenoprotective activity via an increase in endogenous NO generation, suppression of lipid peroxidation and mobilization of antioxidant activity and changes in glycoconjugate content of the gastroduodenal mucosa of rat. [Abstract/Link to Full Text]

Dembi?ski A, Warzecha Z, Ceranowicz P, Dembi?ski M, Cieszkowski J, Pawlik WW, Konturek SJ, Tomaszewska R, H?adki W, Konturek PC
Cannabinoids in acute gastric damage and pancreatitis.
J Physiol Pharmacol. 2006 Nov;57 Suppl 5137-54.
Recent studies have shown that stimulation of cannabinoid 1 (CB1) receptor reduces the area of ischemic myocardial necrosis and affects activity of the digestive tract. The aim of the present study was to check whether the administration of CB1 receptor agonist or antagonist affects the stress-induced gastric ulceration and development of edematous pancreatitis. METHODS: Experiments were performed on rats. Gastric lesions were induced by water immersion and restrain stress (WRS). Acute pancreatitis was induced by cerulein. Prior to WRS or before and during cerulein administration, a natural endogenous ligand for CB1 receptor, anandamide was administered intraperitoneally at the dose of 0.8, 1.5 or 3.0 micromol/kg. A synthetic CB1 receptor antagonist, AM 251 (ALEXIS(R) Biochemicals) was administrated at the dose of 4 micromol/kg i.p. alone or in combination with anandamide at the dose of 1.5 micromol/kg. RESULTS: Administration of anandamide reduced gastric lesions and this effect was associated with am increase in gastric mucosal blood flow and mucosal DNA synthesis; whereas serum level of pro-inflammatory interleukin-1 beta was reduced. Treatment with AM 251 aggravated gastric damage and reversed protective effect of anandamide administration. Opposite effect was observed in the pancreas. Administration of anandamide increased dose-dependently the severity of pancreatitis. In histological examination, we observed an increase in pancreatic edema and inflammatory infiltration. Also, treatment with anandamide augmented the pancreatitis-induced increase in serum level of lipase, amylase, poly-C ribonuclease, and pro-inflammatory interleukin-1 beta; whereas pancreatic DNA synthesis was reduced. Treatment with AM 251 reduced histological and biochemical signs of pancreatic damage and reversed deleterious effect of anandamide in cerulein-induced acute pancreatitis. CONCLUSIONS: Activation of CB1 receptors evokes opposite effects in the stomach and pancreas: in the stomach, exhibits protective effect against stress-induced gastric mucosal lesions; whereas in the pancreas, increases the severity of cerulein-induced pancreatitis. [Abstract/Link to Full Text]

Konturek PC, Kania J, Hahn EG, Konturek JW
Ascorbic acid attenuates aspirin-induced gastric damage: role of inducible nitric oxide synthase.
J Physiol Pharmacol. 2006 Nov;57 Suppl 5125-36.
Aspirin (ASA) represents an important risk factor for gastric mucosal injury. Recently, vitamin C releasing aspirin (ASA-VitC) has been shown to reduce gastric toxicity of ASA in animal model of gastric injury. The aim of the present study was to compare the effect of ASA and ASA-VitC on the gastric mucosal damage before and after Helicobacter pylori (Hp) eradication in 10 young healthy Hp-positive volunteers. All subjects underwent endoscopy at day 0 (before ASA or ASA-VitC treatment) and at day 3 following treatment (1.6 g ASA/day or 1.6 g ASA + 0.96 g Vit C/day). In addition, in vitro experiments were performed in which gastric mucosal cell line (MKN-45 cells) was incubated with ASA or ASA-VitC alone or in combination with H.pylori. Expression of constitutive and inducible NO synthase (cNOS, iNOS) was analyzed by Western blot. Moreover, COX-2 expression was analyzed in gastric biopsies at mRNA and protein level by RT-PCR and Western blot, respectively. In humans, treatment with ASA-VitC induced significantly less gastric mucosal lesions than plain ASA. Furthermore, in comparison to plain ASA, ASA-VitC caused stronger inhibition of cNOS and increase in iNOS expression in the gastric mucosa. In vitro studies demonstrated a significant increase in iNOS expression in MKN-45 cells incubated with Hp. This effect was aggravated by the addition of ASA, but not ASA-VitC, to MKN-45 cells incubated with H.pylori. Both ASA and ASA-VitC stimulated the COX-2 expression in the gastric mucosa. We conclude that ASA-VitC in comparison with ASA induces less gastric mucosal damage and this protective effect may be due to its inhibitory effect on iNOS expression. [Abstract/Link to Full Text]

Botting RM
Inhibitors of cyclooxygenases: mechanisms, selectivity and uses.
J Physiol Pharmacol. 2006 Nov;57 Suppl 5113-24.
The prostaglandins are lipid mediators, discovered in the 1930s by von Euler in Sweden and Goldblatt in the United Kingdom. They are made by the bifunctional enzyme, cyclooxygenase, which has both cyclooxygenase and peroxidase activities in the same molecule. Prostaglandins are involved in physiological functions such as protection of the stomach mucosa, aggregation of platelets and regulation of kidney function. They also have pathological functions such as their involvement in inflammation, fever and pain. Vane in 1971 elegantly showed that the pharmacological actions of aspirin and similar drugs were due to the inhibition of cyclooxygenase. Thus, aspirin-like drugs exert their anti-inflammatory, antipyretic and analgesic effects by inhibition of cyclooxygenase. In 1991, Simmons and his colleagues identified a second cyclooxygenase enzyme, designated cyclooxygenase-2, derived from a separate gene from cyclooxygenase-1. Cyclooxygenase-2 is upregulated by inflammatory mediators and forms prostaglandins which intensify the inflammatory response. Cyclooxygenase-1 is, therefore, a 'housekeeping' enzyme making prostaglandins, which are important for maintaining physiological functions and cyclooxygenase-2 makes prostaglandins which are important in inflammation. The discovery of cyclooxygenase-2 and the establishment of its structure led to the development of selective inhibitors of this enzyme, such as celecoxib and rofecoxib, with potent anti-inflammatory actions but with reduced gastrotoxic effects. A putative cyclooxygenase-3, has also been characterised and cloned. This enzyme is a product of the cyclooxygenase-1 gene, but retains intron 1 after transcription and translates into a cyclooxygenase enzyme with 34 additional amino acids. It is more sensitive to inhibition by paracetamol, aspirin and some other non-steroid anti-inflammatory drugs than cyclooxygenase-1 or cyclooxygenase-2. A cyclooxygenase enzyme induced in cultured cells by some non-steroid anti-inflammatory drugs is also more sensitive to inhibition by paracetamol than cyclooxygenase-2 induced by bacterial lipopolysaccharide. [Abstract/Link to Full Text]

Kotunia A, Zabielski R
Ghrelin in the postnatal development of the gastrointestinal tract.
J Physiol Pharmacol. 2006 Nov;57 Suppl 597-111.
Ghrelin is a 28-amino acid peptide first isolated from rat and human stomachs. Together with the recently discovered 23-amino acid obestatin, it is derived from proghrelin by posttranslational processing. Cells immunoreactive to ghrelin are widely distributed in the gastric mucosa in domestic and laboratory animals and in humans. Ghrelin plays an important role in energy homeostasis, body weight control, and food intake, whereas obestatin seems to induce the opposite effects. Ghrelin and ghrelin receptor expression have been found in developing gastrointestinal foetal and neonatal tissues, and substantial amounts of ghrelin are present in colostrum, thereby suggesting its presumable role in perinatal development. Ghrelin was shown to positively influence weight gain, increase GH, insulin and cortisol secretion. It also stimulates gastrointestinal tissue structure and function development in weaned animals. Surprisingly, responses in suckling neonates were found to be opposite to those in weanlings. Ghrelin retarded gastric, intestinal and pancreatic development and showed a tendency to reduce body weight gain. Recent studies suggest that the biphasic effect of ghrelin in young rats on pancreas and stomach growth seems to be related to age-dependent changes of the release of anabolic IGF-1. In the perinatal period, obestatin is detected in the rat stomach, pancreas and blood plasma. The obestatin concentration in rats is abruptly reduced after birth, contrasting with an increase in the concentration of acylated ghrelin. Further progressive reduction in pancreas obestatin is observed until weaning. [Abstract/Link to Full Text]

Recent Articles in Polish Journal of Pharmacology and Pharmacy

Szelag A, Magdalan J, Kopacz M, Ku?niar A, Kowalski P, Pie?niewska M
Assessment of efficacy of quercetin-5'-sulfonic acid sodium salt in the treatment of acute chromium poisoning: experimental studies.
Pol J Pharmacol. 2003 Nov-Dec;55(6):1097-103.
Hexavalent chromium compounds exhibit higher toxicity than its trivalent compounds since chromium ions in the +6 oxidation state easily cross biological membranes. It has recently been proposed that substances reducing chromium ions from the +6 to the less toxic +3 oxidation state can be beneficial in management of acute chromium poisoning. In vitro studies also demonstrated quercetin-5 '-sulfonic acid sodium salt (NaQSA) to reduce chromium ions from the +6 to the +3 oxidation state. The aim of the study was to determine efficacy of NaQSA in treatment of acute poisoning with a hexavalent chromium compound. The experiment was carried out on male and female Wistar rats which were divided into 4 experimental (A,B,C,D) and control (K) groups. All animals received intragastrically a single CrO3 dose equal to its LD50. Thirty minutes after administration of CrO3, NaQSA was administered intragastrically at a dose of 50 mg/kg (group A) and 100 mg/kg (group B). In groups C and D, NaQSA was administered ip 2 h after administration of CrO3 and then twice a day for 4 days at doses of 50 mg/kg (group C) and 100 mg/kg (group D). Only intragastric administration of NaQSA at a dose of 100 mg/kg decreased mortality in acute poisoning with CrO3. In groups B and D, aminotransferase activity was statistically significantly dropping from day 7 of the experiment in comparison with the group K, which indicates lesser damage to the liver in animals treated with NaQSA. Bilirubin concentrations in groups B and D were also much lower than in the group K, but the difference between average bilirubin levels in these groups and the K was not statistically significant. The results of the study suggest the usefulness of NaQSA in the treatment of poisoning with hexavalent chromium compounds. [Abstract/Link to Full Text]

Wollny T, Chabielska E, Malinowska-Zaprza?ka M, Nazarko J, Rozmys?owicz-Szermi?ska W, Buczko W
Effects of Bulgarian red and white wines on primary hemostasis and experimental thrombosis in rats.
Pol J Pharmacol. 2003 Nov-Dec;55(6):1089-96.
A number of epidemiological studies have demonstrated that moderate red wine consumption significantly decreases the risk of ischemic heart disease. Our earlier studies provide evidence that Italian red wine modulates primary hemostasis and prevents experimental venous thrombosis in rats, independently of its alcohol content, by a nitric oxide (NO)-mediated mechanism. In the present study, we have tested whether Bulgarian red and white wines can influence thrombotic process and primary hemostasis in rats. NO and PGI2 were evaluated as possible mediators of these effects. We have found that red wine treatment (for 10 days) induced a marked prolongation of bleeding time, decrease in platelet adhesion to fibrillar collagen, reduction in venous thrombus weight and shortening of occlusion time in arterial thrombosis model. The fall in venous thrombus weight was also observed after white wine supplementation. Red wine affects hemostasis and venous thrombosis after its iv injection 15 min before experiment. These effects were prevented by NO inhibitor (L-NAME) and PGI2 inhibitor (indomethacin). Our results demonstrate the ability of Bulgarian wines to modulate primary hemostasis and prevent venous and arterial thrombosis in rat. [Abstract/Link to Full Text]

Zieba R, Wagrowska-Danilewicz M
Influence of carnosine on the cardiotoxicity of doxorubicin in rabbits.
Pol J Pharmacol. 2003 Nov-Dec;55(6):1079-87.
The aim of this study was to establish the effect of naturally occurring antioxidant carnosine (CAR) on the doxorubicin (DOX)-induced cardiotoxicity in a rabbit model. For this purpose, we evaluated the influence of DOX administration alone and in a combined therapy with CAR on the hemodynamic parameters and on the degree of cardiac muscle cell alterations in rabbits. Thirty one chinchilla rabbits were divided into four groups. One group of rabbits was injected iv with DOX at a dose of 2 mg kg(-1) weekly for 7 weeks to induce congestive heart failure. Another group of rabbits received the same doses of DOX simultaneously with CAR at a dose of 100 mg kg(-1) po daily for 9 weeks. Administration of CAR started 1 week prior to the first dose of DOX and ended one week after the administration of the last dose of DOX. The control groups of animals received 0.9% NaCl and CAR alone. The following hemodynamic parameters were estimated: heart rate (HR), mean arterial pressure (MAP), cardiac index (CI), stroke index (SI) and total peripheral resistance (TPR). Registration of the hemodynamic parameters in rabbits was performed by Doppler method (Hugo Sachs Elektronik Haemodyn). CAR normalized the values of MAP in rabbits receiving DOX and increased the values of CI and SI. The influence of CAR on TPR was not statistically significant, but there was a decreasing tendency. The degree of cardiac muscle cell alterations was examined by light microscopy using Mean Total Score (MTS) technique. The histopathological studies revealed smaller damage of cardiac muscle in rabbits which received DOX with CAR in comparison to animals receiving DOX alone. CAR seems to be cardioprotective during DOX administration. [Abstract/Link to Full Text]

Bartu? JB, Ch?opicki S
Effect of neutral endopeptidase inhibition on vascular response induced by exogenous angiotensin I in the isolated rat lung.
Pol J Pharmacol. 2003 Nov-Dec;55(6):1071-8.
It is suggested that vasoconstriction mediated by angiotensin II cleaved from angiotensin I by angiotensin converting enzyme (ACE) is counterbalanced by concomitant formation of vasodilator angiotensin (1-7) by neutral endopeptidase (NEP). Here, we tested this hypothesis using as a bioassay the isolated rat lung perfused with Krebs-Henseleit (KH) solution and ventilated with negative pressures. Addition of angiotensin I (100 nM) into the isolated lung resulted in an immediate increase in pulmonary arterial pressure (Delta PAP) which was not accompanied by a significant change in respiratory lung function or weight of the lung. The Delta PAP response induced by angiotensin I was abolished by an inhibitor of ACE, perindoprilate (1 microM), or by angiotensin type 1 receptor antagonist (losartan, 1 microM) but not by angiotensin type 2 receptor antagonist (PD 123.319, 10 microM) suggesting the involvement of ACE and AT1 (but not AT2) receptors in this response. On the other hand, antagonist of bradykinin receptor B2 (icatibant, 100 nM) or an inhibitor of neutral endopeptidase, thiorphan (1 microM and 10 microM) did not modify DeltaPAP response induced by angiotensin I. In summary, in the isolated rat lung perfused with KH solution, ACE has a dominant role in the pulmonary conversion of angiotensin I to angiotensin II, while NEP-derived angiotensin 1-7 does not seem to constitute a major counterbalancing mechanism in the pulmonary vasoconstriction induced by endogenously formed angiotensin II. [Abstract/Link to Full Text]

Hrynyk R, Storm G, Metselaar B, Langner M
Pharmacokinetics of liposomes designed to carry glucocorticoids.
Pol J Pharmacol. 2003 Nov-Dec;55(6):1063-70.
Over the past decade, particulate drug formulations have been successfully employed to reduce undesired side effects and improve drug biodistribution. Despite numerous experimental data, there are relatively few theoretical studies regarding the pharmacokinetics of such formulations. A quantitative pharmacokinetic description of particulate drug forms requires serious adjustments in existing theoretical approaches, due to formulation size. Thus, blood vessel permeabilization and the immunological system need to be accounted for. In this paper, we present a pharmacokinetic model intended to describe the distribution of glucocorticoid (prednisolone phosphate) encapsulated in long-circulated liposomes and its qualitative analysis. In order to achieve qualitative and quantitative agreement with experimental patterns of time-dependent liposome concentration changes in blood, liver and spleen, the existence of two hypothetical liposome populations was assumed. The two populations differ in their accumulation capacities, dosage and time constants. The first population is accumulated in the liver with a time constant of 50 s(-1) and a saturation level of 0.005 micromol/animal, whereas the second with 0.003 s(-1) and 50 micromol/animal, respectively. Such liposome parameterization results from the theoretical model used, however, it may have a physiological foundation. If the two opsonin and/or macrophage types that interact with the liposomes are assumed to have different characteristics, then the pharmacokinetic data obtained experimentally in an animal model can be described correctly. [Abstract/Link to Full Text]

Daniel WA, Kot M, Wójcikowski J
Influence of classic and atypical neuroleptics on caffeine oxidation in rat liver microsomes.
Pol J Pharmacol. 2003 Nov-Dec;55(6):1055-61.
Caffeine is a marker drug for testing the activity of CYP1A2 (3-N-demethylation) in humans and rats. Moreover, CYP3A seems to be essential for its metabolism (8-hydroxylation). In the case of 1-N- and, in particular, 7-N-demethylation of caffeine, apart from CYP1A2, other CYP isoenzymes play a considerable role, probably CYP2B and/or CYP2E1. The aim of the present study was to investigate the influence of two classic neuroleptics (promazine and haloperidol) and two atypical ones (risperidone and sertindole) on cytochrome P-450 activity measured by caffeine oxidation in rat liver microsomes. The obtained results showed that promazine, a phenothiazine neuroleptic with the simplest chemical structure, significantly inhibited 1-N- and 3-N-demethylation and 8-hydroxylation of caffeine via competitive or mixed mechanism (Ki = 21.8, 25.4 and 58.2 microM, respectively). This indicates inhibition by promazine of CYP1A2 (inhibition of 3-N- and 1-N-demethylation), and possibly CYP3A2 (inhibition of 8-hydroxylation), but not of other CYP isoenzymes involved in 7-N-demethylation of caffeine (e.g. CYP2B2 and/or CYP2E1). In contrast to promazine, haloperidol had no effect on the oxidation reactions of caffeine in the applied in vitro metabolic model. The potency of inhibition of caffeine oxidation by risperidone and sertindole resembled rather haloperidol than promazine. Risperidone appeared to be a very weak inhibitor of 3-N-demethylation and 8-hydroxylation (Ki = 202.5 microM) and had no effect on 1-N- and 7-N-demethylation of caffeine. Sertindole was a very poor inhibitor of 1-N- and 7-N-demethylations and 8-hydroxylation pathways of the marker substance (Ki = 132.1, 434.1 and 173.3 microM, respectively); even the observed in vitro inhibition of 3-N-demethylation of caffeine by sertindole (Ki = 68.9 microM) cannot be of practical significance in vivo, considering extremely low pharmacological and therapeutic doses of the neuroleptic. In summary, among the investigated neuroleptics, only promazine showed significant inhibitory activity towards caffeine metabolism in vitro (inhibition of CYP1A2 and possibly CYP3A), which may be of pharmacological and clinical importance in vivo. In contrast to promazine, haloperidol and the investigated atypical neuroleptics had no or very weak effect on caffeine oxidation in vitro,of no in vivo significance. Considering the results of the present and previous studies, it seems highly likely that promazine may cause pharmacokinetic interactions, while atypical neuroleptics seem to be safe in this respect. Moreover, the observed reaction-dependent effects of promazine and sertindole provide indirect evidence that CYP1A2 is not the only isoenzyme important for the metabolism of caffeine, which requires further pharmacological and clinical consideration. [Abstract/Link to Full Text]

Daniel WA, Kot M, Wójcikowski J
Effects of classic and newer antidepressants on the oxidation pathways of caffeine in rat liver. In vitro study.
Pol J Pharmacol. 2003 Nov-Dec;55(6):1045-53.
Caffeine undergoes 3-N-demethylation via CYP1A2, as well as 1-N-demethylation, 7-N-demethylation and 8-hydroxylation, which may involve other CYP isoenzymes. The aim of the present study was to investigate the influence of clomipramine, desipramine, sertraline, nefazodone and mirtazapine on cytochrome P-450 activity measured by caffeine oxidation in rat liver microsomes. The obtained results showed that all the investigated antidepressants, with an exception of mirtazapine, added in vitro to liver microsomes had an inhibitory effect on caffeine metabolism (via competitive or mixed mechanism), though their potency towards particular metabolic pathways was different. Dixon analysis of caffeine metabolism carried out in the control liver microsomes, in the absence and presence of the antidepressant drugs showed that desipramine and clomipramine exerted the most potent inhibitory effect on caffeine metabolism. Desipramine decreased the rates of 1-N-, 3-N- and 7-N-demethylations, and 8-hydroxylation of caffeine (Ki = 23.3, 36.6, 23.3 and 63.3 microM, respectively), the effect on 1-N- and 7-N-demethylation being the most pronounced. Clomipramine showed distinct inibition of 1-N- and 3-N-demethylation and 8-hydroxylation of caffeine, the effects on N-demethylations being the most pronounced (Ki = 38.6, 34.8, 45.6 microM, respectively). Its effect on 7-N-demethylation was rather weak (Ki = 97.8 microM). Sertraline decreased significantly the rate of 1-N- and 3-N-demethylation and 8-hydroxylation (Ki = 37.3, 69.3 and 64 microM, respectively), while its effect on 7-N-demethylation of caffeine was less pronounced (Ki = 92.1 microM). Nefazodone displayed clear effect on 3-N- and 7-N-demethylation (Ki = 68.8 and 66.4 microM, respectively), but was weak in inhibiting 1-N-demethylation and 8-hydroxylation of caffeine (Ki = 110 and 186 microM, respectively). In contrast to the above-tested antidepressants, mirtazapine did not decrease significantly the oxidation rates of 3-N-demethylation or 8-hydroxylation (Ki = 264 and 455 microM, respectively) and had no effect on other oxidation pathways of caffeine. In summary, we have observed intra- and inter-drug differences in the inhibitory effects of the antidepressants on the four oxidation pathways of caffeine in rat liver microsomes. The tested antidepressants (with an exception of mirtazapine) may lead to drug-drug metabolic interactions at a level of a few CYP isoforms. The obtained results provide further indirect evidence that apart from CYP1A2, other CYP isoforms are also important for the metabolism of caffeine. [Abstract/Link to Full Text]

Mahalakshmi K, Pushpakiran G, Anuradha CV
Taurine prevents acrylonitrile-induced oxidative stress in rat brain.
Pol J Pharmacol. 2003 Nov-Dec;55(6):1037-43.
Acrylonitrile (ACN) is a volatile, toxic liquid used as a monomer in the manufacture of synthetic rubber, styrene plastics, acrylic fiber and adhesives. ACN is a potent neurotoxin and a carcinogen, which produces tumors in rats, particularly gliomas of the brain. A role for free radical-mediated lipid peroxidation in the toxicity of ACN has been suggested. We examined the ability of taurine, an antioxidant amino acid, to attenuate ACN-induced alterations in lipid peroxidation, cellular DNA fragmentation, GSH, vitamin C and vitamin E levels in blood and brain of rats. Rats were administered with ACN at a concentration of 100 ppm in drinking water and sacrificed after 14 and 28 days. The level of lipid peroxidation and the enzymatic and non-enzymatic antioxidants were assayed. The obtained data were compared with those obtained from ACN rats co-treated with taurine for 14 and 28 days. It was observed that taurine treatment counteracted the oxidative stress induced by ACN by reducing the levels of peroxidation, and enhancing the activities of enzymatic and non-enzymatic antioxidants. [Abstract/Link to Full Text]

Lena PJ, Subramanian P
Evaluation of the antiperoxidative effects of melatonin in ammonium acetate-treated Wistar rats.
Pol J Pharmacol. 2003 Nov-Dec;55(6):1031-6.
The efficacy of melatonin (MLT) against ammonium acetate-induced neurotoxicity was biochemically studied in the experimental rats. The activities of serum transaminases and the levels of thiobarbituric acid reactive substances were significantly increased in ammonium acetate-treated rats. These levels were significantly decreased in MLT and ammonium acetate-treated rats. Further, non-enzymatic (vitamin C and E) and enzymatic (superoxide dismutase and catalase) antioxidants were significantly decreased in ammonium acetate-treated rats and were increased in MLT and ammonium acetate-treated rats. These biochemical alterations during MLT treatment could be due to its ability to: (i) scavenge a variety of radicals and reactive species, (ii) induce antioxidative enzymes which reduce steady state levels of reactive species, (iii) inhibit nitric oxide synthase which generates nitric oxide and (iv) stabilize cell membranes which assists them in reducing oxidative damage and, thus, prevents the oxidative stress in rats. [Abstract/Link to Full Text]

Veeravalli KK, Akula A, Kota MK
Nitric oxide- and prostaglandin-mediated cardioprotection by bradykinin in myocardial ischemia and reperfusion injury.
Pol J Pharmacol. 2003 Nov-Dec;55(6):1021-9.
The aim of the present study was to investigate the involvement of nitric oxide and prostaglandin pathways in the cardioprotective actions mediated by bradykinin via the combined inhibition of angiotensin converting enzyme and aminopeptidase P in an in vivo rat model of acute ischemia (30 min) and reperfusion (4 h). Myocardial infarction was produced by occlusion of the left anterior descending coronary artery for 30 min followed by 4 h of reperfusion. Infarct size was measured by using the staining agent TTC (2,3,5-triphenyl-tetrazolium chloride). Lipid peroxide levels in serum and in heart tissue were estimated spectrophotometrically. A lead II ECG was monitored at various intervals throughout the experiment. Infarct size expressed as percent of left ventricle was found to be 50.5 +/- 3.5 in control animals and was reduced to 19.4 +/- 1.1 and 15.0 +/- 2.1 with the combined treatment of enalapril or lisinopril and 2-mercaptoethanol, respectively. There was no significant difference in the infarct size of control animals and in the animals treated with HOE140 prior to the combined treatment. Infarct size reduction obtained with the combined inhibition with enalapril and 2-mercaptoethanol or lisinopril and 2-mercaptoethanol was blocked partially but significantly with the prior administration of L-NAME (Nomega-nitro-L-arginine methyl ester) or aspirin, suggesting the involvement of both nitric oxide and prostaglandin pathways in the cardioprotective actions mediated by bradykinin. [Abstract/Link to Full Text]

Boksa J, Charakchieva-Minol S, Duszy?ska B, Bugno R, K?odzi?ska A, Tatarczy?ska E, Chojnacka-Wójcik E, Bojarski AJ
Synthesis, in vitro and in vivo 5-HT1A/5-HT2A serotonin receptor activity of new hybrid 1,2,3,4-tetrahydro-gamma-carbolines with 1-(2-methoxyphenyl)piperazine moiety.
Pol J Pharmacol. 2003 Nov-Dec;55(6):1013-9.
A series of 15 new 2-H- and 2-substituted 5-[omega-[4-(2-methoxyphenyl)-piperazinyl]-alkyl]-1,2,3,4-tetrahydro-gamma-carboline derivatives were prepared, and their affinity for 5-HT1A and 5-HT2A serotonin receptors was determined. Most of those hybrid compounds were found to bind with high affinity to 5-HT1A sites (Ki < 50 nM; 2d, 3a, 3b, 3d, 3e, 4b, 4d, 4e) and moreover two of them (4d, 4e) were mixed 5-HT1A/5-HT2A ligands. The results of a lower lip retraction test in rats indicated that the 2-acetyl derivative with a dimethylene spacer (2d) had features of a postsynaptic 5-HT1A receptor agonist, whereas its analogues with longer chains (3d and 4d) behaved like antagonists. Both 5-HT2A receptor ligands (4d, 4e) at high doses inhibited the (+/-)-DOI-induced head twitches in mice and were classified as weak antagonists of those receptors. [Abstract/Link to Full Text]

Bujalska M
Effect of cyclooxygenase and NO synthase inhibitors administered centrally on antinociceptive action of acetaminophen (Part II).
Pol J Pharmacol. 2003 Nov-Dec;55(6):1001-11.
As it has been demonstrated in a previous study, both cyclooxygenases (COXs) and nitric oxide synthases (NOSs) participate in the mechanism of acetaminophen (ACETA) action. Results obtained in this study indicate that intrathecal (it) or intracerebroventricular (icv) pretreatment with LG-nitro-L-arginine (L-NO-Arg), a non-selective inhibitor of NOS activity, as well as with 7-nitroindazole (7-NI), a selective nNOS inhibitor, potentiated the antinociceptive activity of subceiling doses of ACETA, but were without effect on the action of supramaximal doses in Randall-Selitto test. Similar effect of L-NO-Arg and 7-NI it was observed in writhing test, whereas L-NO-Arg icv or L-NIL it did not influence the action of ACETA in this model. Indomethacin (IND), an inhibitor preferentially acting on COX-1, as well as nimesulide (NIM) and celecoxib (CECOX), i.e. preferential and selective inhibitor of COX-2, respectively, administered icv almost completely blocked the antinociceptive effect of ACETA in Randall-Selitto method. On the other hand, pretreatment with NSAIDs it initially increased and then attenuated the ACETA antinociception. Yohimbine (YOH), an alpha2-adrenergic receptor antagonist, did not modify the antinociceptive action of ACETA administered alone. However, YOH decreased the nociceptive threshold increased by simultaneously administered IND and ACETA, NIM and ACETA, as well as CECOX and ACETA in Randall-Selitto model. In contrast to the peripheral (sc) application, IND administered centrally (icv or it) did not modify the ACETA antinociception in writhing test. Neither NIM nor CECOX administered sc, it or icv changed the ACETA antinociception in this model. Possible mechanisms and sites of antinociceptive effects of ACETA are discussed. [Abstract/Link to Full Text]

Rogóz Z, Skuza G, Wójcikowski J, Daniel WA
Effects of combined treatment with imipramine and metyrapone in the forced swimming test in rats. Behavioral and pharmacokinetic studies.
Pol J Pharmacol. 2003 Nov-Dec;55(6):993-9.
In the present study, we examined the effects of imipramine (IMI) and metyrapone (MET) given alone or in combination of IMI and MET in the forced swimming test in rats. We also measured pharmacokinetic parameters: the level of IMI and its metabolite, desipramine (DMI), in the rat plasma and brain (1 h after the forced swimming test). The present studies indicate that MET (50 mg/kg) reduced immobility time. Combined treatment with MET (50 mg/kg) and IMI (5 or 10 mg/kg) produced stronger antidepressant-like effect than either of drugs given alone. Sulpiride (dopamine D2/3 antagonist), or WAY 100635 (5-HT1A antagonist) but not prazosin, (alpha1-adrenergic antagonist), at doses ineffective in the forced swimming test, inhibited an antidepressant-like effect induced by co-administration of IMI with MET. The active behaviors in that test did not reflect an increase in general activity, since combined administration of IMI and MET failed to alter the locomotor activity of rats, measured in the open field test. MET elevated the concentrations of IMI and DMI in plasma, and the total drug concentration (IMI + DMI) was doubled by MET. In the brain, MET enhanced the concentration of IMI, but decreased that of DMI, consequently, the brain IMI/DMI ratio increased twice. The total drug concentration in the brain (IMI + DMI) was not changed significantly by MET treatment. The obtained results suggest that dopamine D2/3 and 5-HT1A receptors may contribute to the mechanism of synergistic action of IMI and MET in the forced swimming test in rats, and that pharmacokinetic interaction should not have a substantial impact on the MET-induced potentiation of IMI effect, observed in vivo. These findings may be of particular importance to pharmacotherapy of drug-resistant depression. [Abstract/Link to Full Text]

Plech A, Klimkiewicz T, Maksym B
Effect of L-arginine on memory in rats.
Pol J Pharmacol. 2003 Nov-Dec;55(6):987-92.
Effect of intracerebroventricularly (icv) or subcutaneously (sc) injected L-arginine (L-Arg) on memory was determined using the procedure of passive avoidance test. Moreover, locomotor and exploratory activity was determined in rats in an open field test. We found that either the peripheral (sc) or icv administration of L-Arg significantly prolonged latency time in the passive avoidance test. This effect appeared at 20-100-fold higher doses in comparison to such effect of arginine vasopressin (AVP) observed in our previous study. This memory improving effect was not correlated with the inhibition of locomotor and exploratory activity. The effect of the lower icv dose (10 nmoles) of L-Arg was blocked by L-NAME, a non-selective nitric oxide synthase (NOS) inhibitor. Moreover, the effect of both used doses (10 and 100 nmoles) of L-Arg was also blocked by S-methylisothiourea (Mtu), a selective inhibitor of inducible isoform of NOS. On the other hand, the effect of higher icv dose of L-Arg (100 nmoles) was prevented by 7-nitroindazole (7-NI), an inhibitor of neuronal NOS. We conclude that a uniform effect of L-Arg on memory is mediated by different isoforms of NOS, mainly by neuronal and inducible NOS. [Abstract/Link to Full Text]

Smia?owska M, Wiero?ska JM, Szewczyk B
Neuroprotective effect of NPY on kainate neurotoxicity in the hippocampus.
Pol J Pharmacol. 2003 Nov-Dec;55(6):979-86.
Previous studies showed that neuropeptide Y (NPY) inhibited hippocampal epileptiform activity and that endogenous NPY might have neuroprotective effects. Therefore, in the present study, the effect of NPY microinjected intrahippocampally on the kainate-induced lesion and changes in NPY immunoreactivity (-IR) were investigated in rat hippocampus. Male Wistar rats, chronically cannulated, were unilaterally injected with kainic acid (KA) 2.5 nmol/1 microl, or additionally with NPY (470 pmol/1 microl) 30 min before or 30 min after KA injection, into the CA1 or dentate gyrus (DG) area of the hippocampus. Seven days later, their brains were taken out and analyzed histologically to estimate the lesion extent, and immunohistochemically to assess NPY-IR. It was found that KA induced extensive degeneration of CA pyramidal neurons and NPY-IR interneurons in the injected hippocampus. Simultaneously, NPY immunoreactivity appeared in mossy fibres and some granular cells in the contralateral hippocampus. NPY given 30 min after the KA injection into CA1 region, induced significant diminution of the lesion extent in the CA pyramidal layer (diminution by 61%). No significant effect was found when NPY was given 30 min before KA or when rats were microinjected into the DG area. The obtained results indicate neuroprotective action of NPY in some models of the kainate-induced hippocampal degeneration. [Abstract/Link to Full Text]

Kurt M, Bilge SS, Kukula O, Celik S, Kesim Y
Anxiolytic-like profile of propofol, a general anesthetic, in the plus-maze test in mice.
Pol J Pharmacol. 2003 Nov-Dec;55(6):973-7.
The present study was performed to investigate the effect of propofol on anxiety using the elevated plus-maze test. Groups of mice received propofol (20, 40, 60 mg/kg) or diazepam (2 mg/kg), caffeine (30 mg/kg), L-arginine (100 mg/kg), m-chlorophenylpiperazine (m-CPP, 2.5 mg/kg) and then were placed in an elevated plus-maze that was composed of two opposite closed arms and two opposite open arms. Propofol (20, 40, 60 mg/kg) and diazepam (2 mg/kg) significantly increased the percentage of time spent in the open arms compared to control. Caffeine (30 mg/kg) and m-CPP (2.5 mg/kg) decreased the percentage of time spent in the open arms and these effects were antagonized when propofol (40 mg/kg) was administered before the test. L-arginine (100 mg/kg) has also produced anxiogenic effect and this effect was not prevented by propofol. All drugs used in this study did not significantly change locomotor activity. These results suggest that propofol has anxiolytic effect in plus-maze test. [Abstract/Link to Full Text]

Rizwan AN, Ali A, Dua Y, Pal SN, Pillai KK
Effects of gabapentin and antidepressant drug combinations on convulsions and memory in mice.
Pol J Pharmacol. 2003 Nov-Dec;55(6):965-71.
In epileptic patients, neurobehavioral problems such as cognitive impairment, depression, and psychosocial impairments have been described, which may have a pathological and/or iatrogenic basis. For this reason additional treatment is required, beside antiepileptic drug (AED) therapy, to correct the accompanying neurological deficits. However, the rationale behind use of antidepressants along with antiepileptics has been questioned due to proconvulsant effects of the former. In the present study, the effect of gabapentin (GBP) on seizure score and memory is evaluated when it is given alone and in combination with some antidepressants, such as sertraline (SERTR) and alprazolam (ALP). Pentetrazole (PTZ)-induced convulsion and spontaneous alternation behavior (SAB) models were used to study the anticonvulsant effect and effect on memory, respectively. Results showed that addition of SERT to GBP or ALP resulted in reduction of anticonvulsant efficacy of these drugs. However, the combination of GBP + SERT + ALP was superior as far as effect on seizure severity and memory was concerned. [Abstract/Link to Full Text]

Kostowski W, Krza?cik P
Neonatal 5-hydroxytryptamine depletion induces depressive-like behavior in adult rats.
Pol J Pharmacol. 2003 Nov-Dec;55(6):957-63.
The influence of neonatal serotonergic lesion on adult behavior in locomotor and depression models was studied in male Wistar rats. When 3-day-old rats were injected intracisternally with 5,7-dihydroxytryptamine (5,7-DHT), a marked depletion of brain 5-HT was observed when animals were killed 3 months after the treatment. Brain catecholamine content was generally not changed by neurotoxin treatment. The behavioral consequence of intracisternal 5,7-DHT administration to developing rats consisted in reduction of adult rats' activity in the forced swimming test. Both desipramine, and, to the lesser extent, fluoxetine, reversed 5,7-DHT-induced immobility of animals. [Abstract/Link to Full Text]

Wardas J, Zapa?a M, Lorenc-Koci E
Influence of acute and chronic 1,2,3,4-tetrahydroisoquinoline administration on the expression of proenkephalin mRNA in the rat striatum.
Pol J Pharmacol. 2003 Nov-Dec;55(6):951-6.
Animal studies have shown that a depletion of dopamine or blockade of dopamine D2 receptors in the striatum produces an increase in striatal proenkephalin (PENK) mRNA expression and an increase in GABAergic transmission in the globus pallidus. Therefore, it has been suggested that an enhanced striatal PENK mRNA expression may reflect to some extent an increase in the activity of the GABAergic striatopallidal pathway whose overactivity has been suggested to take place in the course of Parkinson's disease. Therefore, the aim of the study was to investigate the role of 1,2,3,4-tetrahydroisoquinoline (TIQ), an endogenous substance suspected of producing parkinsonism in humans, in the regulation of the activity of GABAergic striatopallidal pathway in rats. TIQ administered acutely at the dose of 100 mg/kg ip increased the PENK mRNA expression in the dorsal part of the striatum at two levels I and II (rostral and central striatum, respectively). No changes were noticed in the ventral part of the striatum. Moreover, TIQ given chronically to rats for 3 weeks did not modify the level of PENK mRNA in any examined part of the striatum. The present results show that the effect of TIQ on the PENK mRNA expression is different from that described for proparkinsonian model neurotoxins (MPTP, 6-OHDA) as well as for typical neuroleptics, such as haloperidol. [Abstract/Link to Full Text]

Senkowska A, Ossowska K
Role of metabotropic glutamate receptors in animal models of Parkinson's disease.
Pol J Pharmacol. 2003 Nov-Dec;55(6):935-50.
The efficacy of the majority of drugs currently used for treatment of Parkinson's disease is insufficient. Moreover, such therapeutics are not devoid of serious side effects. Multiple studies on animal models of parkinsonism have shown that new class of drugs, acting selectively on metabotropic glutamate receptors (mGluRs) might be very promising for the future therapy of Parkinson's disease. This review briefly describes changes in glutamatergic transmission in the neuronal circuitry of the extrapyramidal system that occur in parkinsonian patients, contains background information on structure, function and distribution of mGluRs throughout the basal ganglia and concentrates on discussion of the results obtained from numerous animal model studies aimed to establish potential antiparkinsonian properties of various mGluR ligands. The reviewed literature data indicate that among these compounds group I mGluR antagonists and group II mGluR agonists might be beneficial to the treatment of parkinsonian akinesia and muscle rigidity. [Abstract/Link to Full Text]

Skuza G
Potential antidepressant activity of sigma ligands.
Pol J Pharmacol. 2003 Nov-Dec;55(6):923-34.
Despite many years' studies of antidepressant drugs (ADs), their mechanism of action still remains unclear. Recently, it has been postulated that substances capable of reducing neurotransmission at the NMDA complex may represent a new class of ADs. Since several ADs have a high affinity for sigma receptors, the sigma binding site may be a relevant mechanism in antidepressant action. Moreover, sigma ligands are able to modulate the activity of the central neurotransmitter systems, including noradrenergic, serotonergic, dopaminergic and glutamatergic (NMDA) ones, which are seemingly important for the mechanism of action of known ADs. The existence of at least two different subtypes of sigma receptors, denoted sigma1 and sigma2 is now widely accepted. The selective agonists of both sigma receptor subtypes are available at present. In particular, a potential antidepressant activity of sigma1 receptor agonists has been postulated, since the antidepressive-like actions of these compounds have been shown in animal models. This article reviews the findings related to potential antidepressant activity of new, selective sigma ligands. [Abstract/Link to Full Text]

Zambrzycka A, Caka?a M, Kami?ska M
Transition metal ions significantly decrease phospholipase C activity degrading phosphatidylinositol-4,5-bisphosphate in the brain cortex.
Pol J Pharmacol. 2003 Sep-Oct;55(5):915-7.
Highly reactive transition metals, such as copper and iron play an obligatory role in generating of reactive oxygen species (ROS). Many neurodegenerative diseases including Alzheimer's disease (AD) and Parkinson's disease (PD) show increased accumulation of these metals. Phosphoinositide metabolism is altered in neurodegenerative diseases. In the present study, we examined the effect of CuSO(4) and FeCl(2) on phospholipase C (PLC) activity degrading phosphatidylinositol-4,5-bisphosphate (PIP(2)) and phosphatidylinositol (PI) in synaptic plasma membranes (SPM) from the rat brain cortex. We report that 25 microM CuSO(4) and FeCl(2) decreased PIP(2)-PLC activity by 60% and 75%, respectively. However, both compounds had no effect on PI-PLC activity. These data indicated that exclusively PIP(2)-PLC is sensitive to transition metal ions. We suggest that chelators of these metals may protect brain against alteration of phosphoinositide metabolism and might be beneficial in the treatment of neurodegenerative diseases. [Abstract/Link to Full Text]

Zambrzycka A, Kacprzak M
Apolipoprotein E4 and A beta peptide 1-42 inhibit polyphosphoinositide biosynthesis in rat brain cortex.
Pol J Pharmacol. 2003 Sep-Oct;55(5):911-3.
Polyphosphoinositides synthesized by phosphatidylinositol 4-kinase, EC (PI4K) and phosphatidylinositol 4-phosphate 5-kinase, EC (PIP5K) are involved in cell signaling and cytoskeleton function. In this study, the effect of apolipoprotein E4 (apoE4) and amyloid beta peptide A beta 1-42 on PI4K and PIP5K activity in cortical synaptic plasma membranes from rat brain was investigated. The results indicated that 0.25 microM apoE4 inhibited PI4K and PIP5K by 40% and 30%, respectively, but 25 microM A beta 1-42 decreased exclusively PI4K activity by 20%. Although the mechanism of apoE4 and A beta action is unknown and needs future investigation, this study suggests that free radical-dependent protein oxidation may be involved in alteration of these enzymes. [Abstract/Link to Full Text]

Kosiorek P, Hryniewicz A, Bialuk I, Zawadzka A, Winnicka MM
Cannabinoids alter recognition memory in rats.
Pol J Pharmacol. 2003 Sep-Oct;55(5):903-10.
Cannabinoids are known to attenuate learning and memory in both humans and animals. In rodents, disruptive effect of cannabinoids on memory, reversed by SR 141716, a specific CB(1) receptor antagonist, was shown in behavioral tests based on conditioning. There are no data concerning the influence of cannabinoids on recognition memory. Recently, the improvement of recognition memory in cannabinoid CB(1) receptor knock-out mice was reported. Therefore, the purpose of the present study was to determine whether a stable analogue of endogenous cannabinoid anandamide, R-(+)-methanandamide (0.25 and 2.5 mg/kg, i.p.) and a potent CB(1) receptor agonist, CP 55,940 (0.025 and 0.25 mg/kg i.p.) affect recognition memory in rats evaluated in an object recognition test, based on discrimination between the familiar and a new object presented at 1h interval. Because cannabinoids at the higher doses can produce motor inhibition, the influence of both compounds on psychomotor activity was evaluated in an open field test. CP 55,940 and R-(+)-methanandamide, at both doses given once, 15 min before the learning trial, significantly attenuated recognition memory, measured by the difference in exploration of a new object and a duplicate of the familiar object. Moreover, CP 55,940 at the higher dose significantly attenuated ambulation, and bar approaches, and at both doses also rearings, evaluated in an open field, performed immediately after an object recognition test, while R-(+)-methanandamide at both doses did not alter locomotor and exploratory activity of rats. This is the first evidence that cannabinoids impair recognition memory in rats. [Abstract/Link to Full Text]

Wesierska-Gadek J, Gueorguieva M, Horky M
Dual action of cyclin-dependent kinase inhibitors: induction of cell cycle arrest and apoptosis. A comparison of the effects exerted by roscovitine and cisplatin.
Pol J Pharmacol. 2003 Sep-Oct;55(5):895-902.
Cyclin-dependent kinases (CDKs) have recently raised considerable interest in view of their key role in the regulation of the cell cycle progression. In proliferating cells, distinct CDKs associated with specific cyclins coordinate in an orchestrated way the appropriate transition between different phases of the cell cycle. Mutations and/or aberrant expression of distinct CDKs and their regulatory components lead to uncontrolled proliferation and finally to carcinogenesis. However, in post-mitotic neurons, all CDKs with the exception of CDK5 are silent. CDK5, a proline-directed serine/threonine kinase exhibiting a close structural homology to the mitotic CDKs, binds to p35, the neuron-specific regulatory subunit of CDK5. CDK5 is very abundant in mature neurons and seems to regulate neurotransmitter release through phosphorylation and down-regulation of calcium channel activity. Therefore, the inhibition of CDKs in neurons after oxidative stress and in neurodegenerative disorders has a protective action. Selective CDKs inhibitors were developed as promising drugs for cancer therapy due to their ability to arrest cell cycle progression. The aim of this study was to compare the anti-proliferative effect of roscovitine (ROSC), a potent CDKs inhibitor, with that of cisplatin (CP) on human breast cancer MCF-7 cells. ROSC exerted stronger inhibitory effect on proliferation and cell cycle progression of MCF-7 than CP. Accumulation of G(2)/M arrested cells starting 6 h after onset of ROSC treatment coincided with a strong up-regulation of the p53. Reconstitution with caspase-3 sensitized MCF-7 cells to CP action. It implicates that ROSC inhibits more selectively and efficaciously the proliferation of human breast carcinoma cells. [Abstract/Link to Full Text]

Szemraj J, Kawecka I, Lachowicz L, Zyli?ska L
Non-genomic effect of estradiol on plasma membrane calcium pump activity in vitro.
Pol J Pharmacol. 2003 Sep-Oct;55(5):887-93.
The aim of the presented study was to compare the effect of 17-beta-estradiol on the hydrolytic activity of plasma membrane calcium pump (PMCA) purified from excitable (rat cortical synaptosomes) and non-excitable (human erythrocytes) cells. Both types of cell membranes contained different composition of the PMCA isoforms. To elucidate if the hormone action could depend on structure of PMCA protein, we assayed the hormone effect on Ca(2+)-ATP-ases pretreated for 15 and 40 min with trypsin. The full length and trypsin-treated Ca(2+)-ATP-ases were next incubated with 17-beta-estradiol at a concentration of 10(-9) and 10(-7) M. In addition, stimulation of calcium pumps by naturally existing activator, calmodulin, was tested. The activity of synaptosomal and erythrocyte Ca(2+)-ATP-ases was differently altered in the trypsin-treated samples. At physiologically relevant concentration of estradiol (10(-9) M), a significant enhancement of the activity was observed for synaptosomal Ca(2+)-ATP-ase, and a further increase occurred in the enzyme treated with trypsin for 15 min. The highest activity of erythrocyte calcium pump was induced after 40 min of incubation with protease. Moreover, the potency of the truncated calcium pump to promote ATP hydrolysis was approximately 2-fold elevated in the presence of 17-beta-estradiol. Calmodulin significantly stimulated the Ca(2+)-ATP-ase, but only the erythrocyte enzyme digested with trypsin for 15 min. It may be suggested that PMCA is a target for estradiol, that shows different mechanisms of action depending on isoform compositions and structural features of the enzyme. [Abstract/Link to Full Text]

Sypecka J
Different vulnerability to cytotoxicity and susceptibility to protection of progenitors versus mature oligodendrocytes.
Pol J Pharmacol. 2003 Sep-Oct;55(5):881-5.
Oligodendrocytes are known to be particularly vulnerable to the cytotoxic effect evoked by different neurodegenerative processes, such as ischemic insult, hypoxia, hypoglycemia or autoaggressive immunological attack like SM. They are the neural cells that undergo sophisticated process of maturation characterized by huge changes in cell metabolism and morphology. Small bipolar cells differentiate into multiprocessed mature oligodendrocytes capable of myelinating CNS. A question arises whether there are any differences in their sensitivity to excitotoxic events? To address this problem, the cells of two distinct stages of differentiation, i.e. progenitors (O-2A) and mature, myelinating oligodendrocytes (MBP+) were selected for investigation of the effects of such apoptogenic factors as H(2)O(2) or serum-withdrawal in vitro. Primary cultures obtained from the brain hemispheres of 18 days old Wistar rat embryos served after 10 days for the establishing pure oligodendrocyte culture (the "shake-off" method by McCarthy and de Vellis, 1980). Oligodendrocytes were cultured in DMEM with addition of insulin, transferrin and sodium selenite. Cytotoxic influence of selected apoptotic factors as well as neuroprotective effects of CsA were estimated by immunochemical detection. The obtained data suggest that progenitors and mature cells respond to apoptogenic conditions by activation of different molecular pathways and specific cytoprotective conditions should be worked out for each type of the cells. [Abstract/Link to Full Text]

Micha?owska-Wender G, Losy J, Wender M, Januszkiewicz-Lewandowska D, Nowak J
Effect of immunomodulatory treatment of multiple sclerosis on lymphocyte surface immunomarkers.
Pol J Pharmacol. 2003 Sep-Oct;55(5):877-80.
The aim of this study was to analyze the effect of immunomodulatory treatment of multiple sclerosis (MS) on lymphocyte surface immunomarkers. The special attention was given to TCR alpha/beta, gamma/delta and alpha/beta HLA-DR markers. Peripheral blood was obtained from 39 patients with clinically definite R-R MS, fulfilling the criteria of McDonald et al.[5]. The group of 15 patients was treated with interferon beta-1a (Avonex) intramuscularly once a week. The blood was obtained before and after two years of treatment. The other group of 10 patients was treated every day with 20 mg of glatiramer acetate (Copaxone) intracutaneously. Subsets of lymphocytes were analyzed by the method of flow cytometry, using monoclonal antibodies produced by Ortho Diagnostic System. The relative results were evaluated using Immuno Count II program. The frequency of the studied subsets in MS was markedly different from that in healthy persons. The higher number of CD4, TCR alpha/beta positive cells and higher CD4/CD8 ratio was observed. In comparison to healthy individuals, in MS patients a decreased number of TCR gamma/delta, and alpha/beta HLA-DR was found. After therapy with glatiramer acetate, CD3 and CD8 positive lymphocytes were more frequently observed than before the drug administration. The CD4/CD8 ratio was markedly decreased. The effect of interferon beta-1a treatment was similar as in the previous group, i.e. a slight increase in CD3 and CD8 was noticed after therapy. Despite the differences in action of both immunomodulatory drugs, which was established in several studies, we like to stress some similarity in their effect on CD3, CD8, alpha/beta HLA-DR and gamma/delta HLA-DR immunomarkers frequency in lymphocyte, and on the CD4/CD8 ratio. This may mean that there are some common immunological steps of special importance for the clinical effect in MS. [Abstract/Link to Full Text]

Zawada M, Liwie? I, Pernak M, Januszkiewicz-Lewandowska D, Nowicka-Kujawska K, Rembowska J, Lewandowski K, Hertmanowska H, Wender M, Nowak J
MSRV pol sequence copy number as a potential marker of multiple sclerosis.
Pol J Pharmacol. 2003 Sep-Oct;55(5):869-75.
Multiple sclerosis (MS) is a neurological disease in which demyelination in the brain and spinal cord is observed. The causal influence of bacterial/viral infections and genetic/immune factors in the etiology of multiple sclerosis is suggested. Multiple sclerosis-related retrovirus (MSRV) is one of the potential agents, which can lead to development of the disease. The aim of cytogenetic studies was assessment of MSRV pol sequence copy number in patients with MS compared to normal individuals. Cytogenetic slides with interphase nuclei and extended chromatin fibers were prepared from peripheral blood of 16 patients with MS and 10 healthy individuals. Fluorescence in situ hybridization (FISH) with biotinylated product of polymerase chain reaction was used in order to analyze MSRV pol sequence copy number in the examined material. Detection of MSRV pol probe was carried out by immunological reaction with avidin-fluorescein and biotinylated anti-avidin. MSRV pol sequence copy number was significantly greater in MS patients than in normal individuals. Using FISH technique to extended chromatin fibers, it was observed that MSRV pol exists as tandem repeats on various chromosomes. The increased number of MSRV pol sequence has been found on chromatin fibers of MS patients as compared to healthy controls. [Abstract/Link to Full Text]

Malinowska B, Göthert M, Przegali?ski E, Schlicker E
Report on the German-Polish symposium "Thirty years of cooperation between German and Polish pharmacologists. New perspectives in the Common Europe", Bia?owieza, September 18-21, 2003.
Pol J Pharmacol. 2003 Sep-Oct;55(5):807-10. [Abstract/Link to Full Text]

Recent Articles in Acta Pharmacologica Sinica

Ma B, Huang HH, Chen XY, Sun YM, Lin LH, Zhong DF
Biotransformation of metoprolol by the fungus Cunninghamella blakesleeana.
Acta Pharmacol Sin. 2007 Jul;28(7):1067-74.
AIM: To investigate the biotransformation of metoprolol, a beta1-cardioselective adrenoceptor antagonist, by filamentous fungus, and to compare the parallels between microbial transformation and mammalian metabolism. METHODS: Five strains of Cunninghamella (C elegans AS 3.156, C elegans AS 3.2028, C echinulata AS 3.2004, C blakesleeana AS 3.153 and AS 3.910) were screened for the ability to transform metoprolol. The metabolites of metoprolol produced by C blakesleeana AS 3.153 were separated and assayed by liquid chromatography-tandem mass spectrometry (LC/MS(n)). The major metabolites were isolated by semipreparative HPLC and the structures were identified by a combination of LC/MS(n) and nuclear magnetic resonance analysis. RESULTS: Metoprolol was transformed to 7 metabolites; 2 were identified as new metabolites and 5 were known metabolites in mammals. CONCLUSION: The microbial transformation of metoprolol was similar to the metabolism in mammals. The fungi belonging to Cunninghamella species could be used as complementary models for predicting in vivo metabolism and producing quantities of metabolite references for drugs like metoprolol. [Abstract/Link to Full Text]

Cui Q, Yu JH, Wu JN, Tashiro S, Onodera S, Minami M, Ikejima T
P53-mediated cell cycle arrest and apoptosis through a caspase-3- independent, but caspase-9-dependent pathway in oridonin-treated MCF-7 human breast cancer cells.
Acta Pharmacol Sin. 2007 Jul;28(7):1057-66.
AIM: To study the caspase-3-independent mechanisms in oridonin-induced MCF-7 human breast cancer cell apoptosis in vitro. METHODS: The viability of oridonin-treated MCF-7 cells was measured by MTT (thiazole blue) assay. Apoptotic cells with condensed nuclei were visualized by phase contrast microscopy. Nucleosomal DNA fragmentation was assayed by agarose gel electrophoresis. The apoptotic ratio was determined by lactate dehydrogenase assay. Cell cycle alternation and mitochondrial membrane potential were measured by flow cytometric analysis. Bax, Bcl-2, caspase-3, caspase-9, heat shock protein (Hsp)90, p53, p-p53, p21, Poly (ADP-ribose) polymerase (PARP), and the inhibitor of caspase-activated DNase (ICAD) protein expressions were detected by Western blot analysis. RESULTS: Oridonin inhibited cell growth in a time- and dose-dependent manner. Cell cycle was altered through the upregulation of p53 and p21 protein expressions. Pancaspase inhibitor Z-VAD-fmk and calpain inhibitor II both decreased cell death ratio. Nucleosomal DNA fragmentation and the downregulation of DeltaPhimit were detected in oridonin-induced MCF-7 cell apoptosis, which was involved in a postmitochondrial caspase-9-dependent pathway. Decreased Bcl-2 and Hsp90 expression levels and increased Bax and p21 expression levels were positively correlated with elevated levels of phosphorylated p53 phosphorylation. Moreover, PARP was partially cleaved by calpain rather than by caspase-3. CONCLUSION: DNA damage provoked alternations in the mitochondrial and caspase-9 pathways as well as p53-mediated cell cycle arrest, but was not related to caspase-3 activity in oridonin-induced MCF-7 cells. [Abstract/Link to Full Text]

Jiao Y, Ge CM, Meng QH, Cao JP, Tong J, Fan SJ
Dihydroartemisinin is an inhibitor of ovarian cancer cell growth.
Acta Pharmacol Sin. 2007 Jul;28(7):1045-56.
AIM: To investigate the anticancer activity of dihydroartemisinin (DHA), a derivative of antimalaria drug artemisinin in a panel of human ovarian cancer cell lines. METHODS: Cell growth was determined by the MTT viability assay. Apoptosis and cell cycle progression were evaluated by a DNA fragmentation gel electro-phoresis, flow cytometry assay, and TUNEL assay; protein and mRNA expression were analyzed by Western blotting and RT-PCR assay. RESULTS: Artemisinin and its derivatives, including artesunate, arteether, artemether, arteannuin, and DHA, exhibit anticancer growth activities in human ovarian cancer cells. Among them, DHA is the most effective in inhibiting cell growth. Ovarian cancer cell lines are more sensitive (5-10-fold) to DHA treatment compared to normal ovarian cell lines. DHA at micromolar dose levels exhibits a dose- and time-dependent cytotoxicity in ovarian cancer cell lines. Furthermore, DHA induced apoptosis and G2 cell cycle arrest, accompanied by a decrease of Bcl-xL and Bcl-2 and an increase of Bax and Bad. CONCLUSION: The promising results show for the first time that DHA inhibits the growth of human ovarian cancer cells. The selective inhibition of ovarian cancer cell growth, apoptosis induction, and G2 arrest provide in vitro evidence for further studies of DHA as a possible anticancer drug in the clinical treatment of ovarian cancer. [Abstract/Link to Full Text]

She MR, Li JG, Guo KY, Lin W, Du X, Niu XQ
Requirement of reactive oxygen species generation in apoptosis of leukemia cells induced by 2-methoxyestradiol.
Acta Pharmacol Sin. 2007 Jul;28(7):1037-44.
AIM: To investigate the effects of 2-methoxyestradiol (2-ME) on 2 myeloid leukemia cell lines HL-60 and U937, and to explore its mechanisms. METHODS: Human myeloid leukemia cells HL-60 and U937 were used. Measurement of mitochondrial membrane potential (Dym) was performed using 5,5',6,6'-Tetrachloro-1,1',3,3'- tetraethylbenzimidazolylcarbocyanine iodide ( JC-1). Apoptosis and cellular nitric oxide (NO) were detected by flow cytometry using Annexin V and NO sensor dye. Superoxide anion was measured with a fluorescent plate reader by dihydroethidium (DHE). Cytotoxicity was analyzed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium assay. RESULTS: 2-ME resulted in viability decrease in a dose-dependent manner. 2-ME treatment also generated reactive oxygen species (ROS), including NO and superoxide anions, which resulted in mitochondria damage. 2-ME-induced apoptosis was correlated with an increase in ROS. The quenching of ROS with N-acetyl-L-cysteine protected leukemia cells from 2-ME cytotoxicity and prevented apoptosis induction by 2-ME. Furthermore, the addition of manumycin, a farnesyltransferase inhibitor, significantly enhanced apoptosis induced by 2-ME. CONCLUSION: Cellular ROS generation plays an important role in the cytotoxic effect of 2-ME. It is possible to use ROS generation agents, such as manumycin, to enhance the antileukemic effect. The combination strategy needs further in vivo justification and may have potential clinical application. [Abstract/Link to Full Text]

Li QB, Chen ZC, You Y, Zou P
Small interfering RNA of cyclooxygenase-2 induces growth inhibition and apoptosis independently of Bcl-2 in human myeloma RPMI8226 cells.
Acta Pharmacol Sin. 2007 Jul;28(7):1031-6.
AIM: To investigate the effects of small interfering RNA of cyclooxygenase-2 (COX-2) on the proliferation and apoptosis of human multiple myeloma RPMI8226 cells and its relation with the Bcl-2 family in vitro. METHODS: Transcription and expression of COX-2 in human myeloma RPMI8226 cells were checked by RT-PCR and Western blot analysis, respectively. The COX-2 siRNA fragment targeting exon 5 of COX-2 gene was transfected into the cells with the Amaxa nucleofection technique. The inhibition of cell growth was detected by 3-(4,5-dimethyl-2- thiazolyl)-2,5-diphenyl-2H-tetrazolium (MTT) assay. Apoptosis was estimated by Annexin-V/ propidium iodide double-labeled cytometry and confirmed by terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling assay. Bcl-2 and Bax expression was evaluated by Western blot analysis. RESULTS: The COX-2 siRNA fragment could be successfully transfected into RPMI8226 cells, which resulted in the significant inhibition of transcription and expression of COX-2 in the myeloma cells. Proliferation of the transfected cells was inhibited and apoptosis was induced (6.52%+/-0.32%, 12.53%+/-2.52%, 24.39%+/-3.51% and 36.48%+/-4.96% for 0, 12, 24, and 48 h, respectively) in a time-dependent manner (P<0.01). However, the expression of Bcl-2 and Bax in the RPMI8226 cells had no significant changes after nucleofection. CONCLUSION: COX-2 siRNA transfection can suppress COX-2 expression in human myeloma RPMI8226 cells, which leads to growth inhibition and apoptosis independent of Bcl-2. [Abstract/Link to Full Text]

Peng JL, Zhao YG, Mai JH, Pang WK, Guo W, Chen GM, Mo GY, Rao GR, Xu YH
Non-cytolytic antigen clearance in DNA-vaccinated mice with electroporation.
Acta Pharmacol Sin. 2007 Jul;28(7):1024-30.
AIM: To explore the potential of electroporation (EP)-mediated hepatitis B virus (HBV) DNA vaccination for the treatment of chronic HBV infection. METHODS: BALB/c mice were vaccinated with HBV DNA vaccine encoding for the HBV preS(2)-S antigen, combined with or without EP. HBV surface antigen expression plasmid was administered into mice liver via a hydrodynamic injection to mimic HBV infection. The clearance of antigen in the serum and liver was detected by ELISA assay and immunohistochemical staining. The histopathology of the liver tissues was examined by HE staining and serum alanine aminotransferase assay. RESULTS: The immunogenicity of HBV DNA vaccine encoding for the HBV preS(2)- S antigen can be improved by EP-mediated vaccine delivery. The elicited immune responses can indeed reduce the expression of HBV surface antigen (HBsAg) in hepatocytes of the mouse model that was transfected to express HBsAg using the hydrodynamic injection method. The antigen clearance process did not cause significant toxicity to liver tissue, suggesting a non-cytolytic mechanism. CONCLUSION: The EP-aided DNA vaccination may have potential in mediating viral clearance in chronic hepatitis B patients. [Abstract/Link to Full Text]

Wu YM, Chen LL, Yan J, Zhuang CY, Gu ZY
Effect of Porphyromonas gingivalis PrtC on cytokine expression in ECV304 endothelial cells and its level in subgingival plaques from patients with chronic periodontitis.
Acta Pharmacol Sin. 2007 Jul;28(7):1015-23.
AIM: To investigate the effect of the collagenase gene (prtC) product of Porphyromonas gingivalis on inducing host cells to secrete inflammatory cytokines, and to discuss the correlation between the PrtC level in subgingival plaque samples and clinical parameters. METHODS: A prokaryotic expression system pET32a-prtC-Escheria coli BL21DE3 was constructed. Antigenicity and immunoreactivity of the recombinant PrtC protein (rPrtC) was identified by Western blotting. ELISA was applied to detect interleukin (IL)-1alpha, IL-8, and TNF-alpha levels in supernatants from rPrtC-induced human umbilical vein endothelial cells (HUVEC) originated ECV304 cells. Clinical parameters recorded at baseline and after treatment included bleeding on probing (BOP), probing depth (PD), and attachment loss (AL). ELISA was established to measure the PrtC level in 196 subgingival plaque samples from 49 patients with chronic periodontitis. RESULTS: After coincubation with 1 microg/mL rPrtC for 24 h and with 5 or 10 microg/mL rPrtC for 12 h, the levels of IL-1 alpha, IL-8, and TNF-alpha secreted by the ECV304 cells increased significantly (P<0.05). The PrtC level in the BOP-positive or the > or =5 mm AL or > or = 6 mm PD sites was higher than that in the BOP-negative or the < or =2 mm AL or < or =6 mm PD sites (P<0.05), respectively. Compared with baseline, the PrtC levels in different AL sites or in the < or =6 mm PD pockets decreased remarkably after treatment (P<0.01), but in the BOP-positive or in the > 6 mm PD sites, the PrtC levels changed insignificantly (P>0.05). CONCLUSION: rPrtC is able to directly induce host cells to synthesize and secrete IL-1 alpha, IL-8, and TNF-alpha. The PrtC level in subgingival samples is correlated with BOP, AL, and PD. [Abstract/Link to Full Text]

Gao MQ, Guo SB, Chen XH, Du W, Wang CB
Molecular mechanisms of polypeptide from Chlamys farreri protecting HaCaT cells from apoptosis induced by UVA plus UVB.
Acta Pharmacol Sin. 2007 Jul;28(7):1007-14.
AIM: To investigate the mechanism of polypeptide from Chlamys farreri (PCF) protecting HaCaT cells from apoptosis induced by UVA plus UVB in vitro. METHODS: An apoptotic model of UV irradiation-induced HaCaT cells was established. The 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide assay, agarose gel electrophoresis, biochemical methods, and Western blotting were employed in the study. RESULTS: PCF inhibited the UV irradiation-induced apoptosis of HaCaT cells. PCF strongly reduced the intracellular reactive oxygen species level, enhanced activities of superoxide dismutase and glutathione peroxidase and increased the total anti-oxidative capacity in HaCaT cells following UV irradiation. Furthermore, we found that PCF could inhibit the phosphorylation of c-Jun amino-terminal kinase and the activity of caspase-3 in a concentration-dependent manner. CONCLUSION: PCF protected HaCaT cells from apoptosis induced by UVA plus UVB, mainly through decreasing the intracellular ROS level and increasing the activities of anti-oxidative enzymes to block the ROS-JNK-caspase-3-apoptosis signaling pathway. [Abstract/Link to Full Text]

Liu WL, Chen SJ, Chen Y, Sun LM, Zhang W, Zeng YM, Zhou TH, Si JM
Protective effects of heat shock protein70 induced by geranylgeranylacetone in atrophic gastritis in rats.
Acta Pharmacol Sin. 2007 Jul;28(7):1001-6.
AIM: To investigate the effect of geranylgeranylacetone (GGA) on the progression of atrophic gastritis in rats and its potential mechanism. METHODS: Atrophic gastritis was induced in Sprague-Dawley rats with 0.1% ammonia solution, 60% ethanol, and 20 mmol/L deoxycholic acid for 24 weeks. Accompanied by the induction of atrophic gastritis, 200 mg/kg GGA was administered by oral gavage for 8 weeks (weeks 17-24). The histological changes in gastric mucosa were quantitated by the index of inflammation, the gastric mucosal thickness, and the amount of glands of 1 mm horizontal length in antrum. Endogenous heat shock protein (HSP)70 levels and distribution were determined by immunoblotting and immunohistochemistry in gastric mucosa. RESULTS: GGA alleviated the pathological progression of atrophic gastritis with inflammation relief (inflammation index: 1.40 in the GGA group and 1.65 in the atrophic gastritis group) and glandular restoration (mucosal thickness and quantity of glands: 194.3 microm and 38.7 mm in the GGA group; 123.3 microm and 32.7 mm in the atrophic gastritis group; P<0.05). GGA significantly induced HSP70 synthesis (P<0.05). Moreover, quercetin, an inhibitor of HSP70 expression, aggravated the infiltration of inflammatory cells and glandular loss in the antrum. CONCLUSION: GGA prevented the progression of atrophic gastritis in rats via the induction of HSP70 expression. [Abstract/Link to Full Text]

Ma Z, Qiao WT, Xuan CH, Xie JH, Chen QM, Geng YQ
Detection and analysis of bovine foamy virus infection by an indicator cell line.
Acta Pharmacol Sin. 2007 Jul;28(7):994-1000.
AIM: To determine the infectivity and replication strategy of bovine foamy virus (BFV) in different cultured cells using the BFV indicator cell line (BICL) system. METHODS: BFV infection was induced by the co-culture method or the transient transfection of the infectious BFV plasmid [pCMV (cytomegalovirus) - BFV] clone. The infectivity of BFV was monitored by the percentage of green fluorescent protein-positive cells in the BICL. The effect of reverse transcriptase inhibitor zidovudine (AZT) on BFV replication was also evaluated in the BICL. RESULTS: The titer of BFV in fetal bovine lung cells was 4-5-folds more than that in either 293T or HeLa (Cells from Henrietta lacks) cells using the co-culture method, and in the meantime was significantly higher than that produced by the infectious clone pCMV-BFV in the same cells. AZT had only a minor effect on viral titers when added to cells prior to the virus infection. In contrast, viral titers reduced sharply to the level of the negative control when the virus was produced from cells in the presence of AZT. CONCLUSIONS: BICL can be used for the titration of the BFV viral infection in non-cytopathic condition. In addition, we provide important evidence to show that reverse transcription is essential for BFV replication at a late step of viral infection. [Abstract/Link to Full Text]

Chen XP, Qian H, Wu JJ, Ma XW, Gu ZX, Sun HY, Duan YZ, Jin ZL
Expression of vascular endothelial growth factor in cultured human dental follicle cells and its biological roles.
Acta Pharmacol Sin. 2007 Jul;28(7):985-93.
AIM: To investigate the expression of vascular endothelial growth factor (VEGF) in cultured human dental follicle cells (HDFC), and to examine the roles of VEGF in the proliferation, differentiation, and apoptosis of HDFC in vitro. METHODS: Immunocytochemistry, ELISA, and RT-PCR were used to detect the expression and transcription of VEGF in cultured HDFC. The dose-dependent and the time-course effect of VEGF on cell proliferation and alkaline phosphatase (ALP) activity in cultured HDFC were determined by MTT assay and colorimetric ALP assay, respectively. The effect of specific mitogen-activated protein kinase (MAPK) inhibitors (PD98059 and U0126) on the VEGF-mediated HDFC proliferation was also determined by MTT assay. The effect of VEGF on HDFC apoptosis was measured by flow cytometry. RESULTS: VEGF was transcribed and expressed in cultured HDFC. VEGF at 10-300 microg/L significantly increased HDFC proliferation and ALP activity compared to the control. Following 1, 3, 5, or 7 d of stimulation, VEGF induced a significant increase in HDFC proliferation compared with the corresponding control, while VEGF was effective at increasing ALP activity at the incubation time point of 3, 5, or 7 d. PD98059 and U0126 could attenuate the VEGF-mediated HDFC proliferation. Fewer apoptotic cells were observed in the VEGF-treated groups compared to the controls, although the difference was not statistically significant. CONCLUSION: VEGF is expressed in cultured HDFC, and at a proper concentration range can stimulate HDFC proliferation, induce HDFC to differentiate in a "cementoblast/osteoblast" pathway and protect HDFC from apoptosis. The MAPK signaling pathway might be involved in the VEGF-mediated HDFC proliferation. [Abstract/Link to Full Text]

Xie HY, Xia WL, Zhang CC, Wu LM, Ji HF, Cheng Y, Zheng SS
Evaluation of hepatitis B virus replication and proteomic analysis of HepG2.2.15 cell line after cyclosporine A treatment.
Acta Pharmacol Sin. 2007 Jul;28(7):975-84.
AIM: The effect of cyclosporine A (CsA) on hepatitis B virus (HBV) replication was investigated, and proteomics expression differentiation after CsA treatment was studied in order to provide clues to explore the effect of CsA on HBV replication. METHODS: Methyl thiazolyl tetrazolium (MTT) assay was used to evaluate the cytotoxicity of CsA. The HBV replication level in the HBV genomic DNA transfected HepG2.2.15 cell line was determined by an ELISA analysis of hepatitis B surface antigens (HBsAg) and Hepatitis B e antigens (HBeAg) in culture supernatant, while the intracellular HBV DNA replication level was analyzed by slot blot hybridization. Two-dimensional electrophoresis was used to investigate the alteration of protein expression in HepG2.2.15 after CsA treatment in vitro. The differentially-expressed proteins were identified by Matrix-assisted laser desorption/ionization-time of flight mass spectrometry combined with an online database search. RESULTS: CsA was able to inhibit the expression of HBsAg, HBeAg, and HBV DNA replication in vitro in a dose-dependent manner. A proteomics analysis indicated that the expression of 17 proteins changed significantly in the CsA treatment group compared to the control group. Eleven of the 17 proteins were identified, including the overexpression of eukaryotic translation initiation factors (eIF) 3k, otubain 1, 14.3.3 protein, eIF2-1 alpha, eIF5A, and the tyrosine 3/tryptophan 5-mono-oxygenase activation protein in CsA-treated HepG2.2.15 cells. The downregulation of the ferritin light subunit, erythrocyte cytosolic protein of 51 kDa (ECP-51), stathmin 1/oncoprotein, adenine phosphoribosyl-transferase, and the position of a tumor protein, translationally controlled 1, was shifted, suggesting it had undergone posttranslational modifications. CONCLUSION: Our study identified the inhibitory effect of CsA on HBV replication, and found that a group of proteins may be responsible for this inhibitory effect. [Abstract/Link to Full Text]

Luo DL, Gao J, Fan LL, Tang Y, Zhang YY, Han QD
Receptor subtype involved in alpha 1-adrenergic receptor-mediated Ca2+ signaling in cardiomyocytes.
Acta Pharmacol Sin. 2007 Jul;28(7):968-74.
AIM: The enhancement of intracellular Ca2+ signaling in response to alpha 1-adrenergic receptor (alpha 1-AR) stimulation is an essential signal transduction event in the regulation of cardiac functions, such as cardiac growth, cardiac contraction, and cardiac adaptation to various situations. The present study was intended to determine the role(s) of the alpha 1-AR subtype(s) in mediating this response. METHODS: We evaluated the effects of subtype-specific agonists and antagonists of the alpha 1- AR on the intracellular Ca2+ signaling of neonatal rat ventricular myocytes using a confocal microscope. RESULTS: After being cultured for 48 h, the myocytes exhibited spontaneous local Ca2+ release, sparks, and global Ca2+ transients. The activation of the alpha 1-AR with phenylephrine, a selective agonist of the alpha 1-AR, dose-dependently increased the frequency of Ca2+ transients with an EC50 value of 2.3 micromol/L. Blocking the alpha 1A-AR subtype with 5-methylurapidil (5-Mu) inhibited the stimulatory effect of phenylephrine with an IC(50) value of 6.7 nmol/L. In contrast, blockade of the alpha 1B-AR and alpha 1D-AR subtypes with chloroethylclonidine and BMY 7378, respectively, did not affect the phenylephrine effect. Similarly, the local Ca2+ spark numbers were also increased by the activation of the alpha 1-AR, and this effect could be abolished selectively by 5-Mu. More importantly, A61603, a novel selective alpha 1A-AR agonist, mimicked the effects of phenylephrine, but with more potency (EC(50) value =6.9 nmol/L) in the potentiation of Ca2+ transients, and blockade of the alpha 1A-AR by 5-Mu caused abolishment of its effects. CONCLUSION: These results indicate that alpha 1-adrenergic stimulation of intracellular Ca2+ activity is mediated selectively by the alpha 1A-AR. [Abstract/Link to Full Text]

Duan JJ, Ma JH, Zhang PH, Wang XP, Zou AR, Tu DN
Verapamil blocks HERG channel by the helix residue Y652 and F656 in the S6 transmembrane domain.
Acta Pharmacol Sin. 2007 Jul;28(7):959-67.
AIM: The objectives of this study were to investigate the inhibitory action of verapamil on wild-type(WT) and mutation HERG K+ channel current (I(HERG)), and to determine whether mutations in the S6 region are important for the inhibition of I(HERG) by verapamil. METHODS: HERG channels (WT, Y652A, and F656A) were expressed in oocytes of Xenopus laevis and studied using the 2-electrode voltage- clamp technique. RESULTS: WT HERG is blocked in a concentration-dependent manner by verapamil (half-maximal inhibition concentration [IC(50)]=5.1 micromol/L), and the steady state activation and inactivation parameters are shifted to more negative values. However, mutation to Ala of Y652 and F656 located on the S6 domain produced 16-fold and 20-fold increases in IC(50) for IHERG blockade, respectively. Simultaneously, the steady state activation and inactivation parameters for Y652A are also shifted to more negative values in the presence of the blockers. CONCLUSION: Verapamil preferentially binds to and blocks open HERG channels. Tyr-652 and Phe-656, 2 aromatic amino-acid residues in the inner (S6) helix, are critical in the verapamil-binding site. [Abstract/Link to Full Text]

Gu WH, Yang S, Shi WX, Zhen XC, Jin GZ
Effects of (-)-stepholidine on NMDA receptors: comparison with haloperidol and clozapine.
Acta Pharmacol Sin. 2007 Jul;28(7):953-8.
AIM: To examine whether (-)-stepholidine (SPD) has a direct effect on the N-methyl- D-aspartic acid receptors (NMDAR) containing the NMDA receptor subunits NR2A or NR2B and to compare its effect with those of haloperidol (Hal) and clozapine (Cloz). METHODS: NMDAR was transiently expressed in human embryonic kidney 293 (HEK293) cells. Changes in intracellular calcium concentration ([Ca2+]i) induced by NMDAR activation were monitored with Fura-2 ratio imaging techniques. RESULTS: SPD had no significant effects on either subunit of NMDAR at a concentration of less than 100 micromol/L. Hal selectively inhibited NMDAR containing the NR2B subunit, whereas Cloz inhibited both subunits of NMDAR. Although both Hal and Cloz inhibited NR1a/NR2B receptor-mediated Ca2+ influx, their effects were different. Hal was more potent and had a faster peak effect than Cloz. CONCLUSION: Both Hal and Cloz inhibit NMDAR-mediated function, whereas SPD produced only a little inhibition at a high concentration. Based on our other studies, the modulation of SPD on NMDAR function may be via D1 receptor action underlying an indirect mechanism. [Abstract/Link to Full Text]

Ding Q, Fang SH, Zhou Y, Zhang LH, Zhang WP, Chen Z, Wei EQ
Cysteinyl leukotriene receptor 1 partially mediates brain cryoinjury in mice.
Acta Pharmacol Sin. 2007 Jul;28(7):945-52.
AIM: To determine whether the cysteinyl leukotriene receptor 1 (CysLT1 receptor) modulates brain cryoinjury and whether the CysLT1 receptor antagonist pranlukast exerts a time-dependent protective effect on cryoinjury in mice. METHODS: Brain cryoinjury was induced by applying a liquid nitrogen-cooled metal probe to the surface of the skull for 30 s. Brain lesion, neuron density, and endogenous IgG exudation were observed 24 h after cryoinjury. Transcription and the expression of the CysLT1 receptor were detected by RT-PCR and immunoblotting, and the localization of the receptor protein by double immunofluorescence. RESULTS: The mRNA and protein expressions of the CysLT1 receptor were upregulated in the brain 6-24 h after cryoinjury, and the CysLT1 receptor protein was primarily localized in the neurons, not in the astrocytes or microglia. Pre-injury treatments with multi-doses and a single dose of pranlukast (0.1 mg/kg) attenuated cryoinjury; postinjury single dose (0.1 mg/kg) at 30 min (not 1 h) after cryoinjury was also effective. CONCLUSION: The CysLT1 receptor modulates cryoinjury in mice at least partly, and postinjury treatment with its antagonist pranlukast exerts the protective effect with a therapeutic window of 30 min. [Abstract/Link to Full Text]

Santonastasi M, Wehrens XH
Ryanodine receptors as pharmacological targets for heart disease.
Acta Pharmacol Sin. 2007 Jul;28(7):937-44.
Calcium release from intracellular stores plays an important role in the regulation of muscle contraction and electrical signals that determine the heart rhythm. The ryanodine receptor (RyR) is the major calcium (Ca2+) release channel required for excitation-contraction coupling in the heart. Recent studies have demonstrated that RyR are macromolecular complexes comprising of 4 pore-forming channel subunits, each of which is associated with regulatory subunits. Clinical and experimental studies over the past 5 years have provided compelling evidence that intracellular Ca2+ release channels play a pivotal role in the development of cardiac arrhythmias and heart failure. Changes in the channel regulation and subunit composition are believed to cause diastolic calcium leakage from the sarcoplasmic reticulum, which could trigger arrhythmias and weaken cardiac contractility. Therefore, cardiac RyR have emerged as potential therapeutic targets for the treatment of heart disease. Consequently, there is a strong desire to identify and/or develop novel pharmacological agents that may target these Ca2+ signaling pathways. Pharmacological agents known to modulate RyR in the heart, and their potential application towards the treatment of heart disease are discussed in this review. [Abstract/Link to Full Text]

Gu Y, Stornetta RL
Synaptic plasticity, AMPA-R trafficking, and Ras-MAPK signaling.
Acta Pharmacol Sin. 2007 Jul;28(7):928-36.
Synaptic modification of transmission is a general phenomenon expressed at almost every excitatory synapse in the mammalian brain. Over the last three decades, much has been discovered about the cellular, synaptic, molecular, and signaling mechanisms responsible for controlling synaptic transmission and plasticity. Here, we present a brief review of these mechanisms with emphasis on the current understanding of alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid receptor (AMPA-R) trafficking and Ras-mitogen-activated protein kinase (MAPK) signaling events involved in controlling synaptic transmission. [Abstract/Link to Full Text]

Liang CS
Cardiac sympathetic nerve terminal function in congestive heart failure.
Acta Pharmacol Sin. 2007 Jul;28(7):921-7.
Increased cardiac release of norepinephrine (NE) and depleted cardiac stores of NE are two salient features of the human failing heart. Researches from my laboratory have shown that these changes are accompanied by a functional defect of NE uptake in the cardiac sympathetic nerve terminals. Our studies have shown that the decrease of NE uptake is caused by reduction of NE transporter density in the sympathetic nerve endings, and this change is responsible, at least in part, for the increased myocardial interstitial NE, decreased myocardial adrenoceptor density, and increased myocyte apoptosis in experimental cardiomyopathies. We have also provided evidence in both intact animals and cultured PC12 cells that the decrease of NE transporter is induced by the actions of oxidative metabolites of exogenous NE, involving endoplasmic reticulum stress and impaired N-glycosylation of the NE transporter. This change in the cardiac sympathetic NE uptake function, as demonstrated by [123I] metaiodobenzylguanidine in human studies, may not only serve as an important prognostic variable in patients with congestive heart failure, but also be used as a surrogate for the efficacies of various therapeutic interventions for heart failure. Finally, increasing evidence suggests and further studies are needed to show that the cardiac sympathetic nerve terminal function may be a direct target for pharmacologic treatment of congestive heart failure. [Abstract/Link to Full Text]

Deng WJ, Yang XQ, Liang YJ, Chen LM, Yan YY, Shuai XT, Fu LW
FG020326-loaded nanoparticle with PEG and PDLLA improved pharmacodynamics of reversing multidrug resistance in vitro and in vivo.
Acta Pharmacol Sin. 2007 Jun;28(6):913-20.
AIM: FG020326, a novel imidazole derivative, is a potent multidrug-resistance (MDR) modulator in vitro and in vivo. However, FG020326 is insoluble. PEDLLA-FG020326 is a FG020326-loaded nanoparticle formed with diblock copolymers of poly (ethylene glycol)-block-poly (D,L-lactic acid) (PEG:PDLLA, PEDLLA) that can solubilize FG020326. This work was intended to evaluate the pharmacodynamics of PEDLLA-FG020326 on reversing MDR in vitro and in vivo. METHODS: Cytotoxicity was determined by tetrazolium assay. The intracellular accumulation and efflux of doxorubicin (Dox) were detected by fluorescence spectrophotometry. The function of P-glycoprotein was examined by Rhodamine 123 (Rh123) accumulation detected by flow cytometry. The KBv200 cell xenograft model was established to investigate the effect of PEDLLA-FG020326 on reversing MDR in vivo. RESULTS: PEDLLA-FG020326 and FG020326 exhibited 56.4- and 35.9-fold activity in reversing KBv200 cells to vincristine (VCR) resistance, respectively and 14.98- and 7.64-fold to Dox resistance, respectively. PEDLLA-FG020326 was much stronger than FG020326, resulting in the increase of Dox and Rh123 accumulation and the decrease of intracellular Dox extrusion in KBv200 cells. Importantly, PEDLLA-FG020326 exhibited more powerful activity than FG020326 in enhancing the effect of VCR against KBv200 cell xenografts in nude mice, but did not appear more toxic. CONCLUSION: The pharmacodynamics of FG020326 was improved by incorporating it into a micellar nanoparticle formed with PEG-block-PDLLA copolymers. [Abstract/Link to Full Text]

Zhou JY, Wang XF, Tang FD, Zhou JY, Lu GH, Wang Y, Bian RL
Inhibitory effect of 1,8-cineol (eucalyptol) on Egr-1 expression in lipopolysaccharide-stimulated THP-1 cells.
Acta Pharmacol Sin. 2007 Jun;28(6):908-12.
AIM: To study the effects of 1,8-cineol (eucalyptol) on the expression of early growth response factor-1 (Egr-1) and NF-kappaB in the human monocyte THP-1 cell line stimulated by lipopolysaccharide (LPS). METHODS: The THP-1 cells were incubated with serial doses of 1,8-cineol (1, 10, and 100 mg/L, 30 min) before being stimulated with LPS (1 mg/L, 30 min). The localization of Egr-1 in the THP-1 cells was detected by immunofluorescence and a laser scanning confocal microscope. The expression of Egr-1 in the nuclei and whole cell, and NF-kappaB in the nuclei, were measured by Western blot analysis. RESULTS: When stimulated by LPS, the FITC-labeled Egr-1 was detected mainly in the nuclei. Moreover, the expression of Egr-1 in the whole cell increased markedly compared with the control cells. 1,8-Cineol pretreatment decreased the expression of Egr-1 in both the nuclei and whole cell of the LPS-stimulated THP-1 cells, and this effect was concentration-dependent, but there was no reaction on the expression of NF-kappaB in the nuclei protein in the LPS-stimulated THP-1 cells. CONCLUSION: In a concentration-dependent manner, 1,8-Cineol reduces LPS-induced Egr-1 expression in nuclei and in whole cell of THP-1 cells, but shows no effect on NF-kappaB expression. [Abstract/Link to Full Text]

Yim MH, Shin JW, Son JY, Oh SM, Han SH, Cho JH, Cho CK, Yoo HS, Lee YW, Son CG
Soluble components of Hericium erinaceum induce NK cell activation via production of interleukin-12 in mice splenocytes.
Acta Pharmacol Sin. 2007 Jun;28(6):901-7.
AIM: To investigate the immunoregulatory functions of water extracts of Hericium erinaceum (WEHE) focusing on natural killer (NK) cell-based anticancer activities. METHODS: Mouse splenocytes or purely isolated NK cells were stimulated with 1-100 mg/L WEHE for 24 h followed by co-culture with (51)Cr-labeled Yac-1 cells for 4 h, then NK cell-derived cytolytic activity was measured using a radio-release assay. Neutralizing antibodies against mouse interleukin-12 (IL-12) were added into the WEHE-stimulated splenocytes, thereafter, cytotoxicity was measured to examine the involvement of IL-12. RT-PCR and ELISA analyses were performed to confirm the induction of transcription and the translation of IL-12 and interferon-gamma (IFN-gamma) in the WEHE-treated splenocytes. RESULTS: WEHE enhanced the cytolytic activity of total splenocytes towards Yac-1 cells in a dose-dependent manner. However, this activation was not observed when the NK cells isolated from the splenocytes were treated with WEHE. Furthermore, the treatment with antibodies against IL-12 abolished the effect of WEHE on splenocyte-derived cytolytic activity. RT-PCR and ELISA analyses showed the induction of IL-12 and IFN-gamma in the WEHE-treated splenocytes. CONCLUSION: WEHE indirectly activates the cytolytic ability of NK cells via the induction of IL-12 in total splenocytes, and possibly via other immuno-mediators or cellular components. [Abstract/Link to Full Text]

Zhang LL, He DL, Li X, Li L, Zhu GD, Zhang D, Wang XY
Overexpression of coxsackie and adenovirus receptor inhibit growth of human bladder cancer cell in vitro and in vivo.
Acta Pharmacol Sin. 2007 Jun;28(6):895-900.
AIM: To study the effect of the overexpression of coxsackie and the adenovirus receptor (CAR) on the growth of the human bladder cancer cell in vitro and in vivo. METHODS: A retroviral vector pLXSN-CAR expressing CAR was constructed and confirmed by restriction enzyme mapping. The pLXSN-CAR vector and control vector pLXSN were transfected into the PT67 packaging cell line to generate retrovirus with high titer. The CAR-negative T24 cell was infected with the pLXSN-CAR and the pLXSN retrovirus, respectively. The positive clone cells were selected with G418 for 2 weeks. The expression level of the CAR protein was detected by Western blot assay. T24 cell growth in vitro was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Anchorage-independent growth was measured by soft-agar colony formation assay. In vivo cell growth was determined by a nude mice xenograft model. RESULTS: The pLXSN-CAR vector containing full-length CAR cDNA was successfully constructed. Western blot analysis showed that a 46 kDa specific band was found in pLXSN-CA-transfected T24 cells. MTT assay identified the growth inhibition of T24/pLXSN-CAR cells. The cell colony forming ability of T24/pLXSN-CAR cells was significantly lower than that of T24/pLXSN and parental T24 cells. There was a reduction in the tumor size in the T24/pLXSN-CAR group as compared with that of the T24/pLXSN group and parental T24 group. CONCLUSION: The overexpression of CAR in T24 bladder cancer cells can inhibit cell growth both in vitro and in vivo. [Abstract/Link to Full Text]

Zhang H, Gajate C, Yu LP, Fang YX, Mollinedo F
Mitochondrial-derived ROS in edelfosine-induced apoptosis in yeasts and tumor cells.
Acta Pharmacol Sin. 2007 Jun;28(6):888-94.
AIM: To investigate whether a similar process mediates cytotoxicity of 1-O-octadecyl-2-O-methyl-rac-glycero-3-phosphocholine (ET-18-OCH3, edelfosine) in both yeasts and human tumor cells. METHODS: A modified version of a previously described assay for the intracellular conversion of nitro blue tetrazolium to formazan by superoxide anion was used to measure the generation of reactive oxygen species (ROS). Apoptotic yeast cells were detected using terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay. DNA fragmentation and the generation of ROS were measured by cytofluorimetric analysis in Jurkat cells. RESULTS: Edelfosine induced apoptosis in Saccharomyces cerevisiae, as assessed by TUNEL assay. Meanwhile, edelfosine induced a time- and concentration-dependent generation of ROS in yeasts. Rotenone, an inhibitor of the mitochondrial electron transport chain, prevented ROS generation and apoptosis in response to edelfosine in S cerevisiae. alpha-Tocopherol abrogated the edelfosine-induced generation of intracellular ROS and apoptosis. Edelfosine also induced an increase of ROS in human leukemic cells that preceded apoptosis. The overexpression of Bcl-2 by gene transfer abrogated both ROS generation and apoptosis induced by edelfosine in leukemic cells. Changes in the relative mitochondrial membrane potential were detected in both yeasts and Jurkat cells. CONCLUSION: These results indicate that edelfosine induces apoptosis in yeasts in addition to human tumor cells, and this apoptotic process involves mitochondria, likely through mitochondrial-derived ROS. These data also suggest that yeasts can be used as a suitable cell model in elucidating the antitumor mechanism of action of edelfosine. [Abstract/Link to Full Text]

Cao LQ, Chen XL, Wang Q, Huang XH, Zhen MC, Zhang LJ, Li W, Bi J
Upregulation of PTEN involved in rosiglitazone-induced apoptosis in human hepatocellular carcinoma cells.
Acta Pharmacol Sin. 2007 Jun;28(6):879-87.
AIM: To investigate the effects of rosiglitazone, a peroxisome proliferator-activated receptor gamma (PPARgamma) agonist, on the expression of the phosphatase and tensin homologue deleted on chromosome 10 gene (PTEN) and cell growth in hepatocellular carcinoma cells, as well as the underlying mechanisms of these effects. METHODS: RT-PCR and Western blotting analyses were performed to detect transcription and the expression of PTEN in Hep3B cells treated with rosiglitazone. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay was used to evaluate cell growth. Flow cytometry, DNA fragmentation analysis, caspase enzymatic assay, and Hoechst 33258 staining were used to determine cell apoptosis. Furthermore, small interfering RNA was used to suppress PTEN expression. RESULTS: Rosiglitazone increased the expression of PTEN in a dose- and time-dependent manner through the PPARgamma-dependent signal transduction pathway. PTEN upregulation was concomitant with a decreased level of Akt phosphorylation, subsequently resulting in cell growth inhibition and apoptosis in Hep3B cells. PTEN knockdown dramatically blocked these effects of rosiglitazone. Moreover, the exposure of cells to rosiglitazone activated caspases-9 and -3 during apoptotic proceeding. CONCLUSION: Thus, upregulation of PTEN is involved in the inhibition of cell growth and the induction of cell apoptosis by rosiglitazone, suggesting that rosiglitazone may be useful in liver cancer therapy via apoptosis. [Abstract/Link to Full Text]

Xu SP, Sun GP, Shen YX, Peng WR, Wang H, Wei W
Synergistic effect of combining paeonol and cisplatin on apoptotic induction of human hepatoma cell lines.
Acta Pharmacol Sin. 2007 Jun;28(6):869-78.
AIM: To investigate whether paeonol (Pae) has synergistic effects with cisplatin (CDDP) on the growth-inhibition and apoptosis-induction of human hepatoma cell lines HepG2 and SMMC-7721. METHODS: The cytotoxic effect of drugs was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide assay. The coefficient of drug interaction was used to analyze the nature of drug interactions. Morphological changes were observed by acridine orange fluorescence staining. Cell cycle and the apoptosis rate were detected by flow cytometry. Bcl-2 and Bax expression were assayed by immunohistochemical staining. RESULTS: Pae or CDDP had antiproliferative effect on the 2 cell lines in a dose-dependent manner, with different sensitivities to drugs. More interestingly, a synergistic inhibitory effect on the viability of the 2 cell lines was observed after treatment with a combination of Pae (15.63, 31.25, and 62.5 mg/L) with various concentrations of CDDP. Further study showed typical morphological changes of apoptosis if the cells were exposed to the two agents for 24 h. The apoptotic rate of the cells with combination treatment was significantly higher than that of cells treated with Pae or CDDP alone. The expression of Bcl-2 decreased and that of Bax increased in the treated groups, especially in the combination group, with the ratio of Bcl-2/Bax decreasing correspondingly. Additionally, a combination of Pae with CDDP resulted in a stronger S phase arrest, compared with Pae or CDDP alone. CONCLUSION: Pae, in combination with CDDP, had a significantly synergistic growth-inhibitory and apoptosis-inducing effect on the 2 human hepatoma cell lines, which may be useful in hepatoma treatment. [Abstract/Link to Full Text]

Jin W, Qu LF, Chen Q, Chang XZ, Wu J, Shao ZM
Gene expression pattern in apoptotic QGY-7703 cells induced by homoharringtonine.
Acta Pharmacol Sin. 2007 Jun;28(6):859-68.
AIM: To classify the genes responsible for apoptosis in QGY-7703 cells induced by homoharringtonine (HHT). METHODS: Apoptosis in QGY-7703 cells induced by HHT was demonstrated by DNA fragmentation and morphological observation. cDNA microarray technology was used to detect gene transcription, and the result of microarrays for genes was confirmed by RT-PCR. RESULTS: Seventy-eight individual mRNA were identified and their transcription levels changed significantly. Those genes, of which 68% were upregulated and 32% were downregulated, were partially related to apoptosis. They were mostly oncogenes, tumor suppressors, enzymes, and kinases. CONCLUSION: HHT is a potential drug in the treatment of liver cancer. TGF-beta, TNF, FAS, p38MAPK, and p53 apoptosis signaling pathways were activated during apoptosis in QGY-7703 cells. Such inducible genes may play important roles in apoptosis and deserve to be further studied. [Abstract/Link to Full Text]

Guo QC, Shen JN, Jin S, Wang J, Huang G, Zhang LJ, Huang G, Yin JQ, Zou CY, Li MT
Comparative proteomic analysis of human osteosarcoma and SV40-immortalized normal osteoblastic cell lines.
Acta Pharmacol Sin. 2007 Jun;28(6):850-8.
AIM: Comparative proteomics provide a powerful approach in screening for alterations in protein levels and post-translational modifications that are associated with tumors. In the present study, we aimed to identify candidate biomarkers to distinguish osteosarcoma (OS) cells from normal osteoblastic cells. METHODS: We employed 3 OS cell lines (U2OS, IOR/OS9, and SaOS-2), and used the SV40-immortalized normal osteoblastic cell line (hFOB1.19) as the control. The differential protein levels in OS and osteoblastic cells were identified using 2-D gel electrophoresis followed by matrix-assisted laser desorption/ionization-time of flight mass spectrometry analyses. Two proteins of interest, the levels of which were significantly increased in OS cells, were further characterized by Western blot analyses. RESULTS: Twenty-six proteins were identified, the expression level of which was either significantly increased or decreased in the OS cells as compared to the control cells. The expression level of the activator of 90 kDa shock protein ATPase homolog 1 (AHA1), was enhanced 12.4-, 24.1-, and 23.8-fold in SaOS-2, IOR/OS9, and U2OS cells, respectively, and the level of the stomatin-like protein 2 (SLP-2) was increased by 10.4- and 7.8-fold in IOR/OS9 and U2OS cells, respectively, as compared to normal osteoblastic cells. Those observations were confirmed by Western blot analyses. CONCLUSION: A differential proteomic analysis was successfully used to identify AHA1 and SLP-2 that were significantly overproduced in OS cells as compared to normal osteoblastic cells, suggesting that those proteins among others may be effective biomarker candidates for the identification of OS cells. [Abstract/Link to Full Text]

Kang Y, Liao WM, Yuan ZH, Sheng PY, Zhang LJ, Yuan XW, Lei L
In vitro and in vivo induction of bone formation based on adeno-associated virus-mediated BMP-7 gene therapy using human adipose-derived mesenchymal stem cells.
Acta Pharmacol Sin. 2007 Jun;28(6):839-49.
AIM: To determine whether adeno-associated virus (AAV)-2-mediated, bone morphogenetic protein (BMP)-7-expressing human adipose-derived mesenchymal stem cells (ADMS) cells would induce bone formation in vitro and in vivo. METHODS: ADMS cells were harvested from patients undergoing selective suction-assisted lipectomy and transduced with AAV carrying the human BMP-7 gene. Non-transduced cells and cells transduced with AAV serotype 2 carrying the enhanced green fluorescence protein gene served as controls. ADMS cells were qualitatively assessed for the production of BMP-7 and osteocalcin, and subjected to alkaline phosphatase (ALP) and Chinalizarin staining. A total of 2.5 x 10(6) cells mixed with type I collagen were implanted into the hind limb of severe combined immune-deficient (SCID) mice and subjected to a histological analysis 3 weeks post implantation. RESULTS: Transfection of the ADMS cells achieved an efficiency of 99% at d 7. Transduction with AAV2-BMP-7 induced the expression of BMP-7 until d 56, which was markedly increased by d 7. The cells were positively stained for ALP. Osteocalcin production and matrix mineralization further confirmed that these cells differentiated into osteoblasts and induced bone formation in vitro. A histological examination demonstrated that implantation of BMP-7-expressing ADMS cells could induce new bone formation in vivo. CONCLUSION: The present in vitro and in vivo study demonstrated that human ADMS cells would be a promising source of autologous mesenchymal stem cells for BMP gene therapy and tissue engineering. [Abstract/Link to Full Text]

Yao LJ, Wang JQ, Zhao H, Liu JS, Deng AG
Effect of telmisartan on expression of protein kinase C-alpha in kidneys of diabetic mice.
Acta Pharmacol Sin. 2007 Jun;28(6):829-38.
AIM: To investigate the effects of angiotensin receptor blocker (ARB) telmisartan on the expression and distribution of protein kinase C (PKC)-alpha in the kidneys of diabetic mice. METHODS: Diabetic mice were induced with streptozotocin and a group of them were randomly selected for treatment with telmisartan. After 6 weeks, the expression and localization of PKC-alpha in the renal cortex, and the outer and inner medulla were assessed by immunohistochemistry and semiquantitative Western blotting. In addition, expressions of PKC-alpha, transforming growth factor-beta1 (TGF-beta1), and vascular endothelial growth factor (VEGF) in glomeruli were measured by semiquantitative immunohistochemistry. RESULTS: Diabetic and normal mice showed similar distributions of PKC-alpha in the kidneys. The expression of PKC-alpha was found in glomeruli, epithelial cells of proximal tubules, and medullary-collecting duct, while not in the medullary and cortical thick ascending limb, and was different in the epithelial cells of proximal tubules of diabetic nephropathy (DN) mice, PKC-alpha was mostly translocated from the basement membrane to the apical membrane, whereas it was largely translocated from the apical membrane to the basement membrane in epithelial cells of the inner medullary-collecting duct. Western blotting detected increased expression of PKC-alpha in the renal cortex and outer medulla, but not in the inner medulla of DN mice. Enhanced expressions of PKC-alpha, TGF-beta1, and VEGF were shown in the glomeruli of DN mice, where PKC-alpha exhibited a correlation to VEGF, but no correlation to TGF-beta1. ARB telmisartan attenuated alterations of PKC-alpha as mentioned earlier in the DN mice. CONCLUSION: Our findings suggest that PKC-alpha may play a role in the pathogenesis of DN, and that the nephroprotective effects of ARB telmisartan may be partly associated with its influence on PKC-alpha. [Abstract/Link to Full Text]

Recent Articles in Biological & Pharmaceutical Bulletin

Endo S, Matsunaga T, Nagano M, Abe H, Ishikura S, Imamura Y, Hara A
Characterization of an oligomeric carbonyl reductase of dog liver: its identity with peroxisomal tetrameric carbonyl reductase.
Biol Pharm Bull. 2007 Sep;30(9):1787-91.
Dog liver contains an oligomeric NADPH-dependent carbonyl reductase (CR) with substrate specificity for alkyl phenyl ketones, but its endogenous substrate and primary structure remain unknown. In this study, we examined the molecular weight and substrate specificity of the enzyme purified from dog liver. The enzyme is a ca. 100-kDa tetramer composing of 27-kDa subunit, and reduces all-trans-retinal and alpha-dicarbonyl compounds including isatin, which are substrates for pig peroxisomal tetrameric carbonyl reductase (PTCR). In addition, the dog enzyme resembles pig PTCR in inhibitor sensitivity to flavonoids, myristic acid, lithocholic acid, bromosulfophthalein and flufenamic acid. Furthermore, the amino acid sequence of dog CR determined by protein sequencing and cDNA cloning was 84% identical to that of pig PTCR and had a C-terminal peroxisomal targeting signal type 1, Ser-His-Leu. The immunoprecipitation using the anti-pig PTCR antibody shows that the dog enzyme is a major form of soluble NADPH-dependent all-trans-retinal reductase in dog liver. Thus, dog oligomeric CR is PTCR, and may play a role in retinoid metabolism as a retinal reductase. [Abstract/Link to Full Text]

Takahashi M, Kudaka Y, Okumura N, Hirano A, Banno K, Kaneda T
Determination of plasma tenofovir concentrations using a conventional LC-MS method.
Biol Pharm Bull. 2007 Sep;30(9):1784-6.
The quantification of tenofovir, a nucleoside reverse transcriptase inhibitor prescribed once daily, in human plasma is important due to a recent increase in its use. HPLC, however, can not easily detect and quantify tenofovir because of interfering peaks. Therefore, we developed a rapid and conventional LC-MS method, validated by estimating the precision and accuracy for inter- and intraday analysis in the concentration range of 0.019-1.567 microg/ml. The calibration curve was linear in the described concentration range. Average accuracy ranged from 95.9 to 100.7%. Relative standard deviations of both inter- and intraday assays were less than 11.6%. Recovery of tenofovir was more than 80.2%. This novel method provides a conventional, accurate and precise way to determine tenofovir in human plasma samples. [Abstract/Link to Full Text]

Ishizu K, Sunose N, Yamazaki K, Tsuruo T, Sadahiro S, Makuuchi H, Yamori T
Development and characterization of a model of liver metastasis using human colon cancer HCT-116 cells.
Biol Pharm Bull. 2007 Sep;30(9):1779-83.
In order to develop a model of liver metastasis of human gastrointestinal cancer cells, we examined the potential of 10 human colon and stomach cancer cell lines (HT-29, WiDr, HCT-116, HCT-15, HCC-2998, MKN7, MKN28, MKN45, MKN74 and St-4) to form liver metastases in nude mice. Among the cell lines, HCT-116 cells consistently formed gross liver metastases when injected into the spleens of nude mice. In contrast, other human colon and stomach cancer cells produced little or no liver metastasis. In order to analyze the high metastatic potential of HCT-116 cells, the adhesion potential was compared between HCT-116 cells and the other colon cancer cell lines. HCT-116 cells showed more efficient adhesion to fibronectin (FN) than other cells. Furthermore, FN enhanced haptotaxis of HCT-116 cells, but not of other colon cancer cells. The high adhesion potential to FN and enhanced haptotaxis may contribute, at least in part, to the high metastatic potential of HCT-116. To assess the value of this newly developed model of liver metastasis, we compared the ability of four anticancer drugs (fluorouracil, doxifluridine, paclitaxel and irinotecan) to inhibit the formation of liver metastases. Paclitaxel and irinotecan showed strong inhibition of liver metastasis but fluorouracil and doxifluridine showed only slight inhibition. Therefore, this model of metastasis may be useful for screening anti-liver metastatic reagents. These results indicate that the HCT-116 liver-metastasis model should be useful for analyzing the molecular mechanism of liver metastasis and for evaluating new anti-liver metastatic drugs. [Abstract/Link to Full Text]

Hattori Y, Maitani Y
Low-molecular-weight polyethylenimine enhanced gene transfer by cationic cholesterol-based nanoparticle vector.
Biol Pharm Bull. 2007 Sep;30(9):1773-8.
Both polyethylenimine (PEI) polymers and cationic nanoparticles have been widely used for non-viral DNA transfection. Previously, we reported that cationic nanoparticles composed of cholesteryl-3beta-carboxyamidoethylene-N-hydroxyethylamine and Tween 80 (NP-OH) could deliver plasmid DNA (pDNA) with high transfection efficiency. To increase the transfection activity of NP-OH, we investigated the potential synergism of PEI and NP-OH for the transfection of DNA into human prostate tumor PC-3, human cervices tumor Hela, and human lung adenocarcinoma A549 cells. The transfection efficiency with low-molecular PEI (MW 600) was low, but that with a combination of NP-OH and PEI was higher than with NP-OH alone, being comparable to commercially available lipofectamine 2,000 and lipofectamine LTX, with very low cytotoxicity. Low-molecular weight PEI could not compact pDNA in size, but rather might help to dissociate pDNA from the complex and release pDNA from the endosome to cytoplasm by the proton sponge effect. Therefore, the combination of cationic cholesterol-based nanoparticles and a low-molecular PEI has potential as a non-viral DNA vector for gene delivery. [Abstract/Link to Full Text]

Nemoto E, Ueda H, Akimoto M, Natsume H, Morimoto Y
Ability of poly-L-arginine to enhance drug absorption into aqueous humor and vitreous body after instillation in rabbits.
Biol Pharm Bull. 2007 Sep;30(9):1768-72.
The effect of poly-L-arginine with a molecular weight of 35.5 kDa (PLA) on the ocular absorption of hydrophilic molecules after instillation was examined in rabbits in vivo. FITC-labeled dextran (3.8 kDa, FD-4) and pyridoxamine were used as model hyprophilic molecules for absorption. The potential toxicity of PLA was evaluated by microscopic observation of the cornea, production of TNF-alpha, and the thickness of the corneal epithelia and stroma. The concentration of pyridoxamine and FD-4 in aqueous humor 30 min after a single instillation of a solution of PLA was 29- and 16-fold higher than that without PLA, respectively, but the drug concentrations were not determined in the vitreous body. Repetitive instillation of PLA every 30 min for 150 min achieved 31.1- and 13.3-fold increases in pyridoxamine and FD-4 in aqueous humor, respectively. Furthermore, significant amounts of pyridoxamine and FD-4 were detected in the vitreous body after the repetitive instillation of PLA, even although very little of these drugs was detected in the vitreous body in the control eye without PLA. On the other hand, repetitive instillation of PLA did not induce any alteration of corneal epithelial and stromal thickness, production of TNF-alpha, and disruption of the epithelial and stromal morphologies and neutrophil infiltration. Our findings suggest that PLA may be useful in promoting drug delivery of hydrophilic drugs to the ocular tissues without producing any significant corneal damage and inflammation. [Abstract/Link to Full Text]

Nishida K, Okazaki M, Sakamoto R, Inaoka N, Miyake H, Fumoto S, Nakamura J, Nakashima M, Sasaki H, Kakumoto M, Sakaeda T
Change in pharmacokinetics of model compounds with different elimination processes in rats during hypothermia.
Biol Pharm Bull. 2007 Sep;30(9):1763-7.
We compared the pharmacokinetics of model compounds with different elimination processes between hypothermic and normothermic rats, to obtain basic information concerning drug therapy during hypothermia. Male Wistar rats were anesthetized with sodium pentobarbital and kept at temperatures of 37 degrees C (normothermic group) by heat lamp, and 32 degrees C or 28 degrees C (hypothermic group) by external cooling. We chose phenolsulfonphthalein (PSP), indocyanine green (ICG) and fluorescein isothiocyanate (FITC)-dextran (FD-4, Mw 4400) as model compounds to determine changes in clearance pathways during hypothermia therapy. The plasma concentrations of PSP as biliary, urinary and metabolic elimination type were increased significantly in the hypothermic group (32 degrees C, 28 degrees C) after i.v. administration at a dose of 1 mg, compared to the normothermic group (37 degrees C). Each PSP clearance (bile, urine and metabolites) in the hypothermic group was decreased, suggesting an influence of hypothermia on the active elimination process. The decreasing tendency was marked at a temperature of 28 degrees C. Moreover, the plasma concentrations of ICG as the biliary excretion type after i.v. administration to the hypothermic rats at a dose of 1 mg were higher with more than 50% decrease in the total body clearance compared to normothermic rats. On the other hand, there was almost no difference in the i.v. pharmacokinetics of FD-4 as the urinary excretion type between 37 degrees C and 32 degrees C. However, renal clearance of FD-4 was significantly decreased at a temperature of 28 degrees C. Accordingly, the change in pharmacokinetics of a drug in the hypothermic group could differ with the elimination processes. [Abstract/Link to Full Text]

Nishizawa K, Torii K, Kawasaki A, Katada M, Ito M, Terashita K, Aiso S, Matsuoka M
Antidepressant-like effect of Cordyceps sinensis in the mouse tail suspension test.
Biol Pharm Bull. 2007 Sep;30(9):1758-62.
Cordyceps sinensis (CS) has been known as a component of traditional medicines that elicit various biological effects such as anti-fatigue, immunomodulatory, and hypoglycemic actions. Since it has been well-established that fatigue is closely related to depression, we used the tail suspension test (TST) in mice to examine the antidepressant-like effects of hot water extract (HWCS) and supercritical fluid extract (SCCS) of CS. Immobility time in the TST was reduced by administration of SCCS (2.5-10 ml/kg, p.o.) dose-dependently though it was not reduced by treatment with HWCS (500-2000 mg/kg, p.o.). Neither HWCS nor SCCS altered locomotor activity in the open field test, excluding the possibility that the effect of SCCS is due to activation of locomotion. Pretreatment with prazosin (an adrenoreceptor antagonist) or sulpiride (a dopamine D2 receptor antagonist) reduced the effect of SCCS on the immobility time. In contrast, pretreatment with p-chlorophenylalanine (p-CPA, a serotonin synthesis inhibitor) did not alter the anti-immobility effect of SCCS. The last finding is consistent with an additional observation that SCCS had no effect on head twitch response induced by 5-hydroxy-L-tryptophan in mice. Taken altogether, these results suggest that SCCS may elicit an antidepressant-like effect by affecting the adrenergic and dopaminergic systems, but not by affecting the serotonergic system. [Abstract/Link to Full Text]

Nakagawa M, Ohno T, Maruyama R, Okubo M, Nagatsu A, Inoue M, Tanabe H, Takemura G, Minatoguchi S, Fujiwara H
Sesquiterpene lactone suppresses vascular smooth muscle cell proliferation and migration via inhibition of cell cycle progression.
Biol Pharm Bull. 2007 Sep;30(9):1754-7.
Abnormal vascular smooth muscle cell (VSMC) proliferation and migration are involved in restenosis following percutaneous transluminal angioplasty (PTCA) as well as in the development and progression of atherosclerosis. We investigated the mechanisms underlying the inhibitory effect of the sesquiterpene 3-oxo-5alphaH,8betaH-eudesma-1,4(15),7(11)-trien-8,12-olide (1) on rat VSMC proliferation and migration. VSMCs were isolated from rat aorta, and then the effect of 1 on cell proliferation and migration was examined using methylthiazolyldiphenyl-tetrazolium bromide (MTT) and chemotaxis assays, respectively. Compound 1 had a potent inhibitory effect on fetal calf serum-induced VSMC proliferation. This effect correlated with reduced expression of cyclin D(1). In addition, 1 also inhibited platelet derived growth factor (PDGF)-induced migration of VSMCs. These results indicate that 1 is a promising candidate for additional biological evaluation to further define its potential as an inhibitory modulator of VSMC responses that contribute to restenosis following PTCA and to the development and progression of atherosclerosis. [Abstract/Link to Full Text]

Ma Y, Han H, Eun JS, Kim HC, Kim HC, Hong JT, Oh KW
Sanjoinine A isolated from Zizyphi Spinosi Semen augments pentobarbital-induced sleeping behaviors through the modification of GABA-ergic systems.
Biol Pharm Bull. 2007 Sep;30(9):1748-53.
Zizyphi Spinosi Semen (ZSS) has been widely used for the treatment of insomnia in oriental countries. This experiment was performed to investigate whether sanjoinine A, one of major alkaloid compounds of ZSS, has hypnotic effects and/or enhances pentobarbital-induced sleeping behaviors through the gamma-aminobutyric acid (GABA)-ergic systems. Sanjoinine A itself did not induce sleeping at the higher dose used in this experiment. However, sanjoinine A prolonged sleeping time and reduced the sleeping latency induced by pentobarbital in a dose-dependent manner similar to muscimol, a GABA(A) receptor agonist. Sanjoinine A also increased sleeping rate and sleeping time when administered combined with pentobarbital at a sub-hypnotic dosage and showed synergistic effects with muscimol in potentiating sleeping onset and enhancing sleeping time induced by pentobarbital. In addition, both sanjoinine A and pentobarbital increased chloride influx in primary cultured cerebellar granule cells. Sanjoinine A also showed similar effects with muscimol in potentiating chloride influx inducing effects of low dose pentobarbital. Sanjoinine A decreased GABA(A) receptor alpha-subunit expression and increased gamma-subunit expression, and had no effects on the abundance of beta-subunits in primary cultured cerebellar granule cells, showing different subunit expression from pentobarbital. In addition, we found that sanjoinine A also enhanced expression of glutamic acid decarboxylase (GAD), but pentobarbital did not. In conclusion, sanjoinine A itself does not induce sleeping, but it augments pentobarbital-induced sleeping behaviors through the modification of GABA-ergic systems. [Abstract/Link to Full Text]

Fukushima A, Shii D, Sumi T, Kageyama T, Ueno H
Cryptomeria japonica-induced allergic conjunctivitis in mice.
Biol Pharm Bull. 2007 Sep;30(9):1745-7.
Japanese cedar pollen (Cryptomeria japonica, Cry j) is the most common allergen causing pollinosis in Japan. However, short ragweed pollen is used commonly as the antigen for experimentally-induced allergic conjunctivitis (EC) and Cry j-induced EC in mice has not been published. We actively immunized BALB/c mice with Cry j, and then performed a challenge with eye drops containing Cry j. We evaluated the early phase and late phase reactions in the conjunctiva, using Evans blue dye leakage and eosinophil infiltration, respectively. Significant inhibition of the early phase reaction was observed following pre-challenge with eye drops that block histamine H1 receptor in the conjunctiva. Thus, Cry j-induced EC appears to represent a suitable model for the study of pollinosis in Japan. [Abstract/Link to Full Text]

Bajsa J, Singh K, Nanayakkara D, Duke SO, Rimando AM, Evidente A, Tekwani BL
A survey of synthetic and natural phytotoxic compounds and phytoalexins as potential antimalarial compounds.
Biol Pharm Bull. 2007 Sep;30(9):1740-4.
The apicomplexan parasites pathogens such as Plasmodium spp. possess an apicoplast, a plastid organelle similar to those of plants. The apicoplast has some essential plant-like metabolic pathways and processes, making these parasites susceptible to inhibitors of these functions. The main objective of this paper is to determine if phytotoxins with plastid target sites are more likely to be good antiplasmodial compounds than are those with other modes of action. The antiplasmodial activities of some compounds with established phytotoxic action were determined in vitro on a chloroquine (CQ) sensitive (D6, Sierra Leone) strain of Plasmodium falciparum. In this study, we provide in vitro activities of almost 50 such compounds, as well as a few phytoalexins against P. falciparum. Endothall, anisomycin, and cerulenin had sufficient antiplasmodial action to be considered as new lead antimalarial structures. Some derivatives of fusicoccin possessed markedly improved antiplasmodial action than the parent compound. Our results suggest that phytotoxins with plastid targets may not necessarily be better antiplasmodials than those that act at other molecular sites. The herbicides, phytotoxins and the phytoalexins reported here with significant antiplasmodial activity may be useful probes for identification of new antimalarial drug targets and may also be used as new lead structures for new antiplasmodial drug discovery. [Abstract/Link to Full Text]

Sumanont Y, Murakami Y, Tohda M, Vajragupta O, Watanabe H, Matsumoto K
Effects of manganese complexes of curcumin and diacetylcurcumin on kainic acid-induced neurotoxic responses in the rat hippocampus.
Biol Pharm Bull. 2007 Sep;30(9):1732-9.
This study aimed to investigate the mechanism underlying the protective effects of manganese complexes of curcumin (Cp-Mn) and diacetylcurcumin (DiAc-Cp-Mn) on kainic acid (KA)-induced excitotoxicity in the rat hippocampus. Systemic injection of KA (10 mg/kg, i.p.) caused seizures and increased the expression of neurotoxic markers, immediate early genes [c-jun, cyclooxygenase 2 (COX-2), brain-derived neurotrophic factor (BDNF), and heat shock protein 70 (hsp70)] and a delayed response gene [inducible nitric oxide synthase (iNOS)], which were measured at 6 and 72 h after KA injection, respectively, in the hippocampus. Pretreatment with Cp-Mn (50 mg/kg, i.p.) and DiAc-Cp-Mn (50 mg/kg, i.p.) but not with curcumin (50 mg/kg, i.p.) delayed the onset of KA-induced seizure without affecting the seizure score. KA injection induced c-Fos immunoreactivity in DG, CA1, and CA3 hippocampal regions, the expression of which peaked at 6 h after injection. Cp-Mn and DiAc-Cp-Mn treatment significantly decreased c-Fos expression elicited by KA. Moreover, Cp-Mn and DiAc-Cp-Mn administration suppressed the KA-induced expression of c-jun, COX-2, BDNF, and iNOS mRNA, whereas curcumin attenuated only iNOS mRNA expression. No compounds tested had an effect on KA-induced hsp70 expression. It is therefore likely that in addition to radical scavenging and SOD-like activities, the suppression of potential neuronal injury marker expression by Cp-Mn and DiAc-Cp-Mn, contributes to the neuroprotective activities of these compounds, which are superior to those of curcumin, on KA-induced excitotoxicity in the hippocampus. These results suggest the beneficial effects of Cp-Mn, and DiAc-Cp-Mn on the treatment of excitotoxicity-induced neurodegenerative diseases. [Abstract/Link to Full Text]

Sun XL, Ito H, Masuoka T, Kamei C, Hatano T
Effect of Polygala tenuifolia root extract on scopolamine-induced impairment of rat spatial cognition in an eight-arm radial maze task.
Biol Pharm Bull. 2007 Sep;30(9):1727-31.
The effects of Polygala tenuifolia root fractions and the acyl groups of its constituents on the retrieval process of spatial cognition in rats were studied using an eight-arm radial maze task. Oral administration of a precipitate fraction (PTB) obtained by concentration of the n-BuOH-soluble portion from the extract of the roots significantly decreased the number of total errors (TEs) and that of working memory errors (WMEs) at doses of 100 mg/kg and 200 mg/kg. However, it caused no significant decrease in the number of reference memory errors (RMEs). In addition, the saponin-rich fraction (PTBM) obtained by purification of PTB also showed significant decreases in TEs and WMEs at a dose of 100 mg/kg. Among the cinnamic acid derivatives present as the acyl groups in the P. tenuifolia constituents, sinapic acid (SNPA) significantly decreased TEs and WMEs at doses of 10 to 100 mg/kg. These results indicated that P. tenuifolia extracts, PTB and PTBM, and SNPA had a beneficial effect on the memory impairment induced by dysfunction of the cholinergic system in the brain. The memory improvement in the scopolamine-induced memory impairment seen in the radial maze performance was due to improvement in the short-term memory. A contribution of some constituents other than SNPA to the memory improvement was also suggested. [Abstract/Link to Full Text]

Lee BH, Lee JH, Yoon IS, Lee JH, Choi SH, Shin TJ, Pyo MK, Choi WS, Lee SM, Lim Y, Rhim H, Nah SY
Mutations of arginine 222 in pre-transmembrane domain I of mouse 5-HT(3A) receptor abolish 20(R)- but not 20(S)-ginsenoside Rg(3) inhibition of 5-HT-mediated ion currents.
Biol Pharm Bull. 2007 Sep;30(9):1721-6.
Ginsenosides, active ingredients of Panax ginseng, exist as stereoisomers depending on the position of the hydroxyl group on carbon-20; i.e. 20(R)-ginsenoside and 20(S)-ginsenoside are epimers. We previously investigated the structure-activity relationship of the ginsenoside Rg(3) stereoisomers, 20-R-protopanaxatriol-3-[O-beta-D-glucopyranosyl (1-->2)-beta-glucopyranoside], (20(R)-Rg(3)) and 20-S-protopanaxatriol-3-[O-beta-D-glucopyranosyl (1-->2)-beta-glucopyranoside], (20(S)-Rg(3)) in regulating 5-HT(3A) receptor-mediated ion currents (I(5-HT)) expressed in Xenopus oocytes and found that 20(S)-Rg(3) rather than 20(R)-Rg(3) was more stronger inhibitor of I(5-HT). In the present study, we further investigated the effects of 20(R)-Rg(3) and 20(S)-Rg(3) on mouse 5-HT(3A) receptor channel activity after site-directed mutations of 5-HT(3A) receptor facilitation site, which is located at pre-transmembrane domain I (pre-TM1). 5-HT(3A) receptor was expressed in Xenopus oocytes, and I(5-HT) was measured using two-electrode voltage clamp technique. In wild-type, both 20(R)-Rg(3) and 20(S)-Rg(3) inhibited I(5-HT) with concentration-dependent and reversible manner. Induction of 5-HT(3A) receptor facilitation by point mutations of pre-TM1 amino acid residue R222 to R222A, R222D, R222E or R222T not only decreased EC(50) values for I(5-HT) compared to wild-type but also abolished 20(R)-Rg(3)-induced inhibition of I(5-HT). Those mutations also shifted the IC(50) values by 20(S)-Rg(3) into right direction by 2- to 4-folds compared with wild-type. These results indicate that 5-HT(3A) receptor facilitation differentially affects 20(R)-Rg(3)- and 20(S)-Rg(3)-mediated 5-HT(3A) receptor channel regulation. [Abstract/Link to Full Text]

Matsuda Y, Ohsaka K, Yamamoto H, Natsume K, Hirabayashi S, Kounoike M, Inoue M
Comparison of newly developed inhalation anesthesia system and intraperitoneal anesthesia on the hemodynamic state in mice.
Biol Pharm Bull. 2007 Sep;30(9):1716-20.
KNI-472 is the first anesthetic system for mice and rats to incorporate a ventilator. It consists of a newly developed syringe pump-type vaporizer and gas monitor that can deliver accurate concentrations of anesthetic gas at an extremely low airflow. In this study, we compared the hemodynamic effects of isoflurane anesthesia using KNI-472 and intraperitoneal pentobarbital anesthesia. In the isoflurane anesthetic group, Institute of Cancer Research (ICR) mice were anesthetized with 5% isoflurane, followed by endotracheal intubation. Subsequently, they were ventilated mechanically, and anesthesia was maintained with 2% isoflurane for a 60-min period using KNI-472. In the pentobarbital anesthetic group, the ICR mice were anesthetized by an intraperitoneal injection of sodium pentobarbital (70 mg/kg). In isoflurane anesthesia, the heart rate (HR) and mean blood pressure (MBP) were stable. In contrast, in pentobarbital anesthesia, MBP decreased in the first stage after the initiation of anesthesia, after which it gradually increased. The intra-group variability in the estimated skin blood flow (SBF) was higher in the pentobarbital anesthesia than that in the isoflurane anesthesia. The PaO(2) and PaCO(2) values at 15 min after the initiation of pentobarbital anesthesia revealed hypoxia and hypercapnia compared with isoflurane anesthesia. In this study, isoflurane anesthesia using KNI-472, unlike pentobarbital anesthesia, did not induce changes in MBP, SBF, or blood gases. The changes induced by pentobarbital anesthesia were attributed to a change in the depth of anesthesia with time. These results indicate that inhalation anesthesia using KNI-472 is suitable in research on the hemodynamic state in mice. [Abstract/Link to Full Text]

Ha YM, Chung SW, Song S, Lee H, Suh H, Chung HY
4-(6-Hydroxy-2-naphthyl)-1,3-bezendiol: a potent, new tyrosinase inhibitor.
Biol Pharm Bull. 2007 Sep;30(9):1711-5.
Tyrosinase is a key enzyme for melanin biosynthesis and known to be sensitive to ultraviolet light in the presence of oxygen. Therefore, finding effective tyrosinase inhibitors, either from synthetic or natural sources, can be beneficial in the treatment of melanin-related disorders. We synthesized 4-(6-hydroxy-2-naphthyl)-1,3-bezendiol (HNB), a new family of hydroxyl substituted phenyl naphthalenes, as the isosteres of oxyresveratrol. This study investigated inhibitory effects of HNB on tyrosinase activity. HNB inhibited mushroom tyrosinase with an IC(50) value of 0.07 microM, which is more potent than the anti-tyrosinase activity of kojic acid (IC(50)=38.24), a well-known tyrosinase inhibitor. The kinetic analysis of tyrosinase inhibition revealed that HNB is a competitive inhibitor (K(i) 4.78 x 10(-9) M at 0.125 microM and K(i) 6.21 x 10(-9) M at 0.25 microM). We further found that HNB also inhibited melanin production in B16F10 melanoma cells (B16 cells). In addition to tyrosinase inhibiting activity, melanin biosynthesis was inhibited by HNB in the B16F10 cells. These data strongly suggest that HNB can suppress the production of melanin via the modulation of tyrosinase activity. [Abstract/Link to Full Text]

Jiang S, Nakano Y, Yatsuzuka R, Ono R, Kamei C
Inhibitory effects of Moutan cortex on immediate allergic reactions.
Biol Pharm Bull. 2007 Sep;30(9):1707-10.
The anti-allergic effect of an ethanol extract from Moutan Cortex was evaluated in some animal models. The Moutan Cortex extract (30, 100 mg/kg, i.p.) dose-dependently inhibited systemic anaphylactic shock induced by compound 48/80 in mice. It also inhibited dose-dependently the scratching behavior induced by compound 48/80 or histamine at a dose of 100 mg/kg. An increase in the vascular permeability induced by compound 48/80 or histamine was also inhibited by the Moutan Cortex. In addition, in vitro studies, the Moutan Cortex inhibited histamine release from rat peritoneal mast cells induced by compound 48/80. To investigate the active component of Moutan Cortex extract, it was suspended in water and extracted with EtOAc to yield EtOAc insoluble (A) and soluble (B) fractions. The effect of extract (B) was more potent than that of extract (A) in inhibiting histamine release. From these findings, it seems likely that the Moutan Cortex extract is effective in antagonizing certain pharmacological effects induced by compound 48/80, which is probably mediated by inhibiting the release of histamine from mast cells and antagonizing the effect on histamine. The main active component of Moutan Cortex is considered to be contained in extract (B). In conclusion, Moutan Cortex may be useful for the relief of symptoms of atopic dermatitis and other allergy-related diseases. [Abstract/Link to Full Text]

Wang X, Zhao X, Li D, Lou YQ, Lin ZB, Zhang GL
Effects of Ganoderma lucidum polysaccharide on CYP2E1, CYP1A2 and CYP3A activities in BCG-immune hepatic injury in rats.
Biol Pharm Bull. 2007 Sep;30(9):1702-6.
The purpose of the present study was to investigate the effect of Ganoderma lucidum polysaccharide (GLPS), a major active component in Chinese medicinal fungus, on cytochrome P450 metabolic activity in Bacillus Calmette Guérin (BCG)-induced immune hepatic injury in rats. The enzyme kinetics of the probes including chlorzoxazone (CYP2E1), phenacetin (CYP1A2) and nifedipine (CYP3A) were evaluated by HPLC. The results showed that BCG-pretreatment (125 mg/kg) significantly increased serum levels of alanine transaminase (ALT), nitrite and malondialdehyde (MDA), inhibited activities of superoxide dismutase (SOD) and decreased P450 total content in microsomes (p<0.05). Administration of GLPS (50 and 200 mg/kg) reversed above hepatic injury stimulated by BCG in vivo. Moreover, GLPS dose-dependently inhibited activities of CYP2E1, CYP1A2 and CYP3A in hepatic microsomes in vitro, suggesting that inhibition of GLPS on P450 oxidative metabolism might participate in the hepatoprotective mechanism, and also suggested that pharmacokinetics might be changed by drug-herb interaction. [Abstract/Link to Full Text]

Neri P, Tokoro S, Yokoyama S, Miura T, Murata T, Nishida Y, Kajimoto T, Tsujino S, Inazu T, Usui T, Mori H
Monovalent Gb3-/Gb2-derivatives conjugated with a phosphatidyl residue: a novel class of Shiga toxin-neutralizing agent.
Biol Pharm Bull. 2007 Sep;30(9):1697-701.
Shiga toxin (Stx) exerts toxic activity by binding to glycosphingolipids, mainly globotriaosyl (Gb(3)) ceramide, on the surface of target cells. The inhibition of toxin-receptor binding is a promising therapeutic approach to prevent Stx-mediated diseases. In this study, we synthesized monovalent Stx-ligands of phosphatidylethanolamine dipalmitoyl-Gb(3) (Gb(3)-PEDP) and galabiosyl (Gb(2))-PEDP and we examined their neutralizing activity against Stx-1 and Stx-2 in vitro. Both Gb(3)-PEDP and Gb(2)-PEDP strongly neutralized the cytotoxicity of Stx-1 and Stx-2. It is likely that the mechanism of neutralization involved formation of liposomes and consequently clustering of sugar units. We propose monovalent Gb(3)-/Gb(2)-derivatives conjugated with phosphatidyl residue as a novel class of Stx-neutralizing agent. [Abstract/Link to Full Text]

Han Y
Synergic anticandidal effect of epigallocatechin-O-gallate combined with amphotericin B in a murine model of disseminated candidiasis and its anticandidal mechanism.
Biol Pharm Bull. 2007 Sep;30(9):1693-6.
In the present study, we investigated synergic anticandidal effect of epigallocatechin-O-gallate (EGCG) in a murine model of disseminated candidiasis caused by Candida albicans. In addition, its mechanism was examined. In the animal system, EGCG-given BALB/c mice group intraperitoneally (i.p.) before intravenous (i.v.) inoculation with viable C. albicans yeast cells survived longer than diluent-received (control) mice group (p<0.05). EGCG treatment inhibited the hyphal formation from the yeast form of C. albicans, causing growth-inhibition of the candidal cells. In experiments determining synergic effect, mice given diluent (control), Amp B (amphotericin B; 0.5 mg/kg of body weight), or EGCG (2 mg/kg) had mean survival times (MST) of approximately 10.9, 11.7, and 13.9 d, respectively. However, mice administered combination of Amp B (0.5 mg/kg) plus EGCG (2 mg/kg) had a MST value of 42.1 d, surviving an average of app. 30 d longer than the Amp B alone-received mice groups. The MST value from the combination-treated mice groups was much greater than MST value from mice groups that received four times the Amp B dose. These results indicate that EGCG, which has anticandidal activity causing blockage of the hyphal formation, has the synergism combined with Amp B against disseminated candidiasis. [Abstract/Link to Full Text]

Futami K, Shimamoto A, Furuichi Y
Mitochondrial and nuclear localization of human Pif1 helicase.
Biol Pharm Bull. 2007 Sep;30(9):1685-92.
Yeast Pif1 DNA helicase is the prototype member of a helicase subfamily participating in the maintenance of telomere, ribosome, and mitochondria DNAs. The Pif1 DNA helicase family is highly conserved from yeast to human, but the biochemical nature of human homologues remains to be clarified. To this end, we investigated the transcriptional unit of human Pif1 gene and its encoded protein hPif1. The results showed that the hPif1 gene product has at least two isoforms consisting of the conserved helicase motif and differential C-terminal regions derived from alternative splicing of the gene transcript. Deletion mutant analysis showed that Pif1 helicase has nuclear localization signal and mitochondria targeting signal at the N-terminal and C-terminal regions, respectively. In HeLa cells, hPif1 helicase expression was induced by the release of cells from serum starvation, suggesting that hPif1 has roles in the S phase. Consistently, the down regulation of the hPif1 helicase by RNA interference with siRNA caused a cell cycle delay at the S phase. These findings suggest that hPif1 in the nucleus may be involved in chromosome maintenance in association with DNA replication, while the function of hPif1 remains to be clarified. [Abstract/Link to Full Text]

Azuma Y, Sato H, Higai K, Matsumoto K
Enhanced expression of membrane-associated sialidase Neu3 decreases GD3 and increases GM3 on the surface of Jurkat cells during etoposide-induced apoptosis.
Biol Pharm Bull. 2007 Sep;30(9):1680-4.
We previously reported that, in Jurkat human T cells, the topoisomerase II inhibitor etoposide enhances sialidase activity and reduces cell surface sialic acid levels at an early stage of apoptosis and that the decreases in sialic acid are suppressed by the sialidase inhibitor 2,3-dehydro-2-deoxy-N-acetylneuraminic acid [Azuma Y., et al., Glycoconj. J., 17, 301-306 (2000)]. In the current studies, we treated Jurkat cells with etoposide and examined the changes in the cell surface levels of gangliosides GM1, GM2, GM3, GD1a, and GD3 at physiological pH using anti-ganglioside antibodies. We also examined the sialidase activity on the cell surface using 4-methylumbelliferyl N-acetylneuraminic acid and measured the mRNA expression of the plasma membrane-associated sialidase Neu3 and the lysozomal Neu1 using real-time PCR. We found an increase in GM3 and a decrease in GD3 during the early stage (4 h) of etoposide-induced apoptosis that preceded the increase in cell surface exposure of phosphatidylserine (4 to 6 h). The caspase 3 inhibitor acetyl-Asp-Glu-Val-Asp-aldehyde significantly suppressed changes in GM3 and GD3 and blocked the enhanced cell surface sialidase activity. Furthermore, etoposide caused a gradual up-regulation of Neu3 mRNA expression but not Neu1 mRNA expression. Enhanced Neu3 mRNA expression was suppressed in the presence of caspase 3 inhibitor. These results indicate that Neu3 is up-regulated in Jurkat cells undergoing etoposide-induced apoptosis through intracellular signaling events downstream of caspase 3 activation and that enhanced Neu3 activity is closely related to the changes of cell surface ganglioside composition. [Abstract/Link to Full Text]

Kim YM, Namkoong S, Yun YG, Hong HD, Lee YC, Ha KS, Lee H, Kwon HJ, Kwon YG, Kim YM
Water extract of Korean red ginseng stimulates angiogenesis by activating the PI3K/Akt-dependent ERK1/2 and eNOS pathways in human umbilical vein endothelial cells.
Biol Pharm Bull. 2007 Sep;30(9):1674-9.
Angiogenesis is important for promoting cardiovascular disease, wound healing, and tissue regeneration. We investigated the effects of Korean red ginseng water extract (KRGE) on angiogenesis and its underlying signal mechanism. KRGE increased in vitro proliferation, migration, and tube formation of human umbilical vein endothelial cells, as well as stimulated in vivo angiogenesis without increasing VEGF expression. KRGE-induced angiogenesis was accompanied by phosphorylation of ERK1/2, phosphatidylinositol 3-kinase (Akt), and endothelial nitric oxide synthase (eNOS) as well as an increase in NO production. Inhibition of PI3K activity by wortmannin completely inhibited KRGE-induced angiogenesis and phosphorylation of Akt, ERK1/2, and eNOS, indicating that PI3K/Akt activation is an upstream event of the KRGE-mediated angiogenic pathway. The MEK inhibitor PD98059 blocked KRGE-induced ERK1/2 phosphorylation without affecting Akt and eNOS activation. However, the eNOS inhibitor N(G)-monomethyl-L-arginine effectively inhibited tube formation, but partially blocked proliferation and migration as well as ERK phosphorylation, without altering Akt and eNOS activation, revealing that the eNOS/NO pathway is partially involved in ERK1/2 activation. This study demonstrated that KRGE stimulates in vitro and in vivo angiogenesis through the activation of the PI3K/Akt-dependent ERK1/2 and eNOS signal pathways and their cross talk. [Abstract/Link to Full Text]

Moon TC, Son SY, Chang HW
Purification and characterization of 45 kDa PAF acetylhydrolase from bovine colostrum.
Biol Pharm Bull. 2007 Sep;30(9):1668-73.
Platelet activating factor (1-O-alkyl-2-acetyl-sn-glycero-3-phosphocholine; PAF) acetylhydrolase (PAF-AH) activity has been identified in bovine colostrum and high levels of this activity are found in early colostrum (within 24 h after parturition). In this study, PAF-AH in early colostrum was purified by ammonium sulfate precipitation, and sequential use of butyl-Toyopearl 650M, DEAE-Sepharose, heparin-Sepharose, hydroxyapatite, chelating-Sepharose and Mono Q HPLC column chromatography. This enzyme is a monomeric polypeptide with a molecular weight of approximately 45 kDa on 12.5% SDS-PAGE. The V(max) and K(m) for PAF-AH were 87.6 microM and 7.96 nmol/min/mg respectively. This enzyme was inhibited by phenylmethylsulfonyl fluoride, iodoacetamide and p-bromophenacylbromide, suggesting that both serine and histidine residues are required for enzyme activity. It was not inactivated by NaF or dithiothreitol. The purified enzyme did not degrade phospholipids with a long chain fatty acyl group at the sn-2 position. Accordingly, this enzyme is distinct from phospholipase A(2). In addition, PAF-AH selectively hydrolyzed oxidatively modified phosphatidylcholine. Furthermore, this enzyme was shown by Western blot analysis using antibody to human plasma PAF-AH to be plasma type PAF-AH. These results clearly demonstrate that 45 kDa plasma type PAF-AH activity exists in bovine colostrum. [Abstract/Link to Full Text]

Takeda O, Takechi S, Ito S, Omori H, Katoh T, Yamaguchi T
Effects of phenyl derivatives of dihydropyrazines with ability to generate radical species on Escherichia coli.
Biol Pharm Bull. 2007 Sep;30(9):1663-7.
Phenyl-substituted dihydropyrazines (Ph-DHPs) are derivatives of 2,3-dihydro-5,6-dimethylpyrazine (Me-DHP). Upon the addition of Cu(2+), Me-DHP inhibits the growth of Escherichia coli by generating hydroxyl and carbon-centered radicals that cause DNA strand breakage. Here, we investigated the toxic effect of Ph-DHPs in several DNA repair-deficient or detoxifying enzyme-deficient mutant strains. Ph-DHPs caused cytotoxic and genotoxic damage, but, in a sodA sodB strain, the effects in the presence or absence of Cu(2+) were different than those of Me-DHP. Our results suggest that the action of the generated superoxide anion in the interior side of the cell is remarkable. [Abstract/Link to Full Text]

Yan B, Wang G, A J, Xie L, Hao H, Liang Y, Sun J, Li X, Zheng Y
Construction of the fingerprints of ginseng stem and leaf saponin reference substances and spiked plasma sample by LC-ESI/MS and its application to analyzing the compounds absorbed into blood after oral administration of ginseng stem and leaf saponin in rat.
Biol Pharm Bull. 2007 Sep;30(9):1657-62.
Develop a simple and reliable assay method to detect ginseng stem and leaf saponins (GSLS) in methanol and rat plasma by liquid chromatography-electrospray ionization mass spectrometry in scan mode, and construct the fingerprints of GSLS reference substances and plasma samples. In order to screen the active constituents of GSLS, analysis and comparison were carried out between the LC/ESI-MS profiles of blank rat plasma and rat plasma samples obtained after oral administration of GSLS. Thirty-one compounds were detected and 10 of them were identified in the fingerprints of reference substances and spiked plasma sample. Furthermore, 12 compounds (C7, C8, C14, C15, C18, Re, C24, Rb(1), Rc, Rb(2), Rb(3), Rd) were absorbed easily and some new compounds were generated after oral administration of GSLS, which might be the metabolites of GSLS. These absorbed components and new compounds may be the main bioactive components of GSLS. [Abstract/Link to Full Text]

Mitamura K, Horikawa A, Yamane Y, Ikeda Y, Fujii Y, Shimada K
Determination of digoxin in human serum using stable isotope dilution liquid chromatography/electrospray ionization-tandem mass spectrometry.
Biol Pharm Bull. 2007 Sep;30(9):1653-6.
A method for the determination of digoxin in human serum using a liquid chromatography/electrospray ionization-tandem mass spectrometry (LC/ESI-MS/MS) technique is reported. Digoxin and the internal standard, [21,21,22-(2)H(3)]digoxin, were extracted from 250 mul of human serum using a solid phase extraction cartridge (Oasis HLB) and analyzed by LC/ESI-MS/MS in the selected reaction monitoring mode. The intra- and inter-assay reproducibility and accuracy were satisfactory within the quantification range of 0.20-3.20 ng/ml. The concentrations of digoxin in the serum samples obtained from digitalized patients (n=19) were in the range of 0.25-2.84 ng/ml, which were compared to those obtained by radioimmunoassay. [Abstract/Link to Full Text]

Toyota E, Iyaguchi D, Sekizaki H, Itoh K, Tanizawa K
Kinetic properties of three isoforms of trypsin isolated from the pyloric caeca of chum salmon (Oncorhynchus keta).
Biol Pharm Bull. 2007 Sep;30(9):1648-52.
Three isoforms of anionic chum salmon trypsin (ST-1, ST-2, and ST-3) were purified from the pyloric caeca of chum salmon (Oncorhynchus keta). The molecular weights of the three isoforms were about 24 kDa as determined by SDS-PAGE. The isoelectric points of ST-1, ST-2, and ST-3 were 5.8, 5.4, and 5.6, respectively. The apparent K(m) values of two isoforms (ST-1 and ST-2) for BAPA (benzoyl-L-arginine-p-nitroanilide) hydrolysis at 5, 15, 25 and 35 degrees C were slightly higher than that of the main isoform ST-3, depending on temperature. The turnover numbers, k(cat), of ST-1 and ST-2 were about twice as high as that of ST-3. Consequently, the catalytic efficiencies (k(cat)/K(m)) of ST-1 and ST-2 were more efficient than ST-3. There were marked differences in both apparent K(m) and k(cat) values of three anionic chum salmon trypsins as compared to bovine cationic trypsin. K(m) values of all chum salmon trypsins were approximately 10 times lower than those of bovine trypsin, depending on the temperature. The k(cat) values of all chum salmon trypsins were about 2- to 5-fold higher than those of bovine trypsin; therefore, the catalytic efficiencies (k(cat)/K(m)) of chum salmon trypsin were 20- to 40-fold more efficient than those of bovine trypsin. On the other hand, k(cat)/K(m) values of ST-1 for TAME (tosyl-L-arginine methyl ester) hydrolysis were lower than those of bovine trypsin, whereas k(cat)/K(m) values of ST-2 and ST-3 were comparable to those of bovine trypsin, depending on the temperature. [Abstract/Link to Full Text]

Suzuki S, Nishida S, Ohno K, Santa T
Modulation of coactivator recruitment by cooperative ligand binding to human Estrogen receptor alpha and beta.
Biol Pharm Bull. 2007 Sep;30(9):1641-7.
Estrogen receptor (ER) is a member of the nuclear receptor superfamily, and functions as a ligand-dependent transcription factor with roles in cell growth and differentiation. In addition to endogenous estrogen, 17beta-estradiol (E(2)) and artificial antagonists, many suspected environmental estrogenic chemicals are reported to bind to ER, with various affinities and transcriptional responses. ER is also an allosteric protein and shows a positive cooperative interaction with E(2). Cooperativity affects inter-subunit interaction, and while ligand-bound ER interacts with coactivators, antagonist-bound ER does not. We therefore hypothesized that ligand-binding characteristics influence coactivator recruitment to the ER dimer, and thereby affect transcriptional activity. We investigated the interaction between ER and human Steroid Receptor Coactivator-1 (SRC-1), in the presence of compounds exhibiting various Hill coefficients. In the case of both ER subtypes (ERalpha and ERbeta), the Hill coefficients of the compounds tested correlated with the affinity of the ER-ligand complex to SRC-1, with the exception of ERbeta-4-n-nonylphenol and ER-antagonist complexes. This is the first report to investigate the relationship between Hill coefficients of ligand binding and coactivator interaction with the ER-ligand complex. We also examined the proteolytic digestion of ER using trypsin, in the presence and absence of compounds with various Hill coefficients, to investigate ligand-dependent conformational changes in ER. We used not only agonists and antagonists but also compounds of weak biological activity (partial agonists). Our results shed light on the subtle modulation of transcriptional activation by chemical agents. [Abstract/Link to Full Text]

Kuge Y, Takai N, Ishino S, Temma T, Shiomi M, Saji H
Distribution profiles of membrane Type-1 matrix metalloproteinase (MT1-MMP), matrix metalloproteinase-2 (MMP-2) and cyclooxygenase-2 (COX-2) in rabbit atherosclerosis: comparison with plaque instability analysis.
Biol Pharm Bull. 2007 Sep;30(9):1634-40.
BACKGROUND: Despite increasing evidence that membrane type 1 matrix metalloproteinase (MT1-MMP), matrix metalloproteinase-2 (MMP-2), and cyclooxygenase-2 (COX-2) are involved in the pathogenesis of atherosclerosis, the possible links among these enzymes remain unclear. Accordingly, we investigated the distribution of MT1-MMP, MMP-2, and COX-2 immunohistologically in the atherosclerotic lesions of hypercholesterolemic (WHHLMI) rabbits. METHODS AND RESULTS: Distribution of MT1-MMP, MMP-2, and COX-2 was examined by immunohistochemical staining using sixty cross sections of the ascending-arch and thoracic aortas prepared from 4 WHHLMI rabbits. MT1-MMP and MMP-2 staining was prominently observed in the macrophage-rich regions of the atheromatous lesions, and was positively correlated with morphological vulnerability (r=0.63 for MT1-MMP; r=0.60 for MMP-2; p<0.0001). MT1-MMP staining was positively correlated with MMP-2 staining (r=0.61, p<0.0001). COX-2 staining was also the highest in the macrophage-rich regions of the atheromatous lesions, with relatively high staining levels in other more stable lesions. CONCLUSIONS: Co-distribution of MT1-MMP, MMP-2, and COX-2 was demonstrated in grade IV atheroma, indicating a possible link among these enzymes in the destabilization of atherosclerotic plaques. The relatively high COX-2 distribution in other more stable lesions may indicate its additional roles in the stabilization of atherosclerotic lesions. The present findings in hypercholesterolemic rabbits should help advance our understanding of the pathophysiology of atherosclerosis and provide useful information for the development of new therapeutic and diagnostic (imaging) agents that target MMPs and COX-2 in atherosclerosis. [Abstract/Link to Full Text]

Recent Articles in Chemical & Pharmaceutical Bulletin (Tokyo)

Chen SG, Chen JJ, Gao K
Prenylisoflavone derivatives from the roots of Hedysarum scoparium.
Chem Pharm Bull (Tokyo). 2007 Aug;55(8):1181-4.
Four new prenylisoflavone derivatives, namely, 5-hydroxy-4'-methoxy-8-prenyl-2''-hydroxyisopropyldihydrofurano[4,5:6,7]-isoflavone (1), 5-hydroxy-4'-methoxy-6-prenyl-2'''-hydroxyisopropyldihydrofurano[4,5:8,7]-isoflavone (2), 5-hydroxy-4'-methoxy-8-prenyl-1'',2''-peroxyl-3'',3''-dimethyldihydropyrano[5,6:6,7]-isoflavone (3), and 5-hydroxy-4'-methoxy-6-prenyl-1''',2'''-peroxyl-3''',3'''-dimethyldihydropyrano[5,6:8,7]-isoflavone (4), together with three known ones 5-7, were isolated from the roots of Hedysarum scoparium. Their structures were established by means of detailed spectroscopic analysis (IR, EI- or HR-ESI-MS as well as 1D- and 2D-NMR), and by comparison of their spectroscopic data with those reported for structurally related compounds. [Abstract/Link to Full Text]

Bao S, Ding Y, Deng Z, Proksch P, Lin W
Rhyncosides A-F, phenolic constituents from the Chinese mangrove plant Bruguiera sexangula var. rhynchopetala.
Chem Pharm Bull (Tokyo). 2007 Aug;55(8):1175-80.
Chemical investigation on the stem of a Chinese mangrove plant Bruguiera sexangula var. rhynchopetala (Rhizophoraceae) resulted in the isolation and characterization of four new phenolic glycosides rhyncosides A-D (1-4), and two new lignan derivatives namely rhyncosides E-F (5-6), along with twelve known phenolic constituents. Their structures were determined by extensive spectroscopic data analyses. [Abstract/Link to Full Text]

Hamashita T, Nakagawa Y, Aketo T, Watano S
Granulation of core particles suitable for film coating by agitation fluidized bed I. Optimum formulation for core particles and development of a novel friability test method.
Chem Pharm Bull (Tokyo). 2007 Aug;55(8):1169-74.
To prepare powdered medicines without bitter taste, film coating is required to cover the surface of core particles. In this study, effect of formulation and operating conditions of agitation fluidized bed on the core particle properties was investigated. In order to prevent breakage of the core particles during coating process, which sometimes causes variation of drug dissolution rate, addition of maltose syrup powder during the formulation process of the core particles was investigated. Also, a method for friability test in which the core particles were subjected to strong impact was proposed to evaluate strength of the core particles. The friability of the core particles determined by this test method correlated well with the actual friability of the particles during the coating process. Based on this result, we confirmed this novel friability test method could predict the core particle endurance during the coating process. [Abstract/Link to Full Text]

Takeda C, Takahashi Y, Seto I, Kawano G, Takayama K, Onishi H, Machida Y
Influence of pectins on preparation characteristics of lactoferrin bioadhesive tablets.
Chem Pharm Bull (Tokyo). 2007 Aug;55(8):1164-8.
For the treatment of chronic inflammation in the oral cavity, we attempted to develop bioadhesive tablets of bovine lactoferrin (B-LF). Pectin was used as a bioadhesive polymer, and the influence of the degree of esterification and the molecular weight of pectins on the characteristics of B-LF tablets were investigated. Concerning bioadhesive force, a tendency increasing the value according to increase of the esterification of the pectin was confirmed. Sustained release of B-LF from the tablets was observed as the esterification increased, and a possibility for prediction of the time required to release 50% of B-LF by using the equation given by the degree of esterification and the logarithm of the molecular weight was suggested. Pectin cross-linked with Ca(2+) (Ca-PC) was also used for the preparation of the B-LF tablets. Prolonged release of B-LF from the tablets was observed as the Ca(2+) in Ca-PC increased. Our findings suggest that pectin with a high degree of esterification is suitable as a bioadhesive polymer since high bioadhesive force and sustained release are shown. Furthermore, a possibility that the B-LF release could be controlled by adjusting the Ca(2+) concentration in Ca-PC was suggested. [Abstract/Link to Full Text]

Jiménez MM, Fresno MJ, Ramírez A
Rheological study of binary gels with Carbopol Ultrez 10 and hyaluronic acid.
Chem Pharm Bull (Tokyo). 2007 Aug;55(8):1157-63.
The objective of this study is to provide a rheological characterization of binary hydroalcoholic gels made with Carbopol Ultrez 10 (U10) and Hyaluronic Acid (HA) as a function of polymer concentration: U10 (0.0-2.0% w/w) and HA (0.00-0.20% w/w), and to determine the influence of this combination on the thixotropic properties of the resulting binary systems. Interaction of the two polymers was measured using the Viscose Synergy Index (I(S)) and thixotropic analysis, which indicate the structural changes that take place in binary gels attributable to molecular interactions between the gelling agents. The maximum values for viscose synergy (I(S)=1.22-1.44) are obtained for the U10 : HA mixtures with a polymer proportion of 10 : 1. The behavior of the binary gels studied is the result of the formation of a more structured three-dimensional network between the U10 and HA molecules. Shearing of this polymer network requires application of a greater force than is needed to shear the structure of the separate gels. Inclusion of HA in a proportion of 1 : 10 has a fixing effect on the polymer network, resulting in greater resistance to shearing in the compound gel. The relative thixotropic area -A(R)- shows maximum values (A(R)=17.215%) for the same polymer composition. The evolution of the two parameters indicates that restructuring of the molecular interactions for this polymer proportion (10 : 1) takes place; the result is a reinforced three-dimensional structure in the gelled system, which increases the thixotropic properties. The same composition leads to a maximum of thixotropic properties as well as viscose synergy because both characteristics are closely related to structural changes observed in the binary systems of this composition. Thixotropic systems have a very wide area of application in the pharmaceutical industry. For this reason, the results obtained here considerably increase the use of the gels studied. In fact, incorporation of HA significantly improves a property of acrylic gels which has direct repercussions on the ease and efficiency of their application to the skin. [Abstract/Link to Full Text]

Lee JH, Kiyota N, Ikeda T, Nohara T
Acyclic diterpene glycosides, capsianosides C, D, E, F and III, from the fruits of hot red pepper Capsicum annuum L. used in Kimchi and their revised structures.
Chem Pharm Bull (Tokyo). 2007 Aug;55(8):1151-6.
Acyclic diterpene glycosides, named capsianosides I', II, III (1), C (2), D (3), E (4) and F (5), have been isolated from the dried hot red pepper fruits of Capsicum annuum L. used in Kimchi. The structures of these compounds have been revised in the sugar connectivities by 1D- and 2D-NMR spectroscopic and chemical methods. [Abstract/Link to Full Text]

Zhou WW, Lin WH, Guo SX
Two new polyporusterones isolated from the sclerotia of Polyporus umbellatus.
Chem Pharm Bull (Tokyo). 2007 Aug;55(8):1148-50.
In the course of searching for marker components, two new polyporusterones were isolated from the sclerotia of Polyporus umbellatus, together with another three known analogs. The structures of the new ones were elucidated as (20S,22R,24R)-16,22-epoxy-3beta,14alpha,23beta,25-tetrahydroxyergost-7-en-6-one (1) and (23R,24R,25R)-23,26-epoxy-3beta,14alpha,21alpha,22alpha-tetrahydroxyergost-7-en-6-one (2) by chemical and spectroscopic means, including HR-FAB-MS, 1D- and 2D-NMR. [Abstract/Link to Full Text]

Musial W
The effect of methylcellulose on metronidazole release from polyacrylic acid hydrogels.
Chem Pharm Bull (Tokyo). 2007 Aug;55(8):1141-7.
Topical treatment of acne rosacea, a chronic condition characterized by recurrent course for many years, is primarily based on metronidazole preparations. The aim of this study was to evaluate the effect of various acrylic acid polymers, in composition with methylcellulose on metronidazole release rate from hydrogels proposed for the treatment of acne rosacea. Viscosity and release studies using "Paddle over Disk" system with semipermeable membrane of MWCO 3500 were performed. Compositions of Carbopol 971P and methylcellulose revealed an increase in viscosity with increasing concentration of methylcellulose in the range of 17200-26166 mPa.s. In all the examined formulations, the release process was characterized by a two-stage course. Among bipolymeric formulations, the highest first-stage release rate of 9.18 x 10(-3) min(-1) was determined for the gel consisting of 2.00% Carbopol 980NF with 1.00% methylcellulose. The second-stage release rates ranged between 2.88 x 10(-3) and 8.00 x 10(-3) min(-1). Two-stage release course can thus be attributed to metronidazole distribution into two compartments of hydrogel matrix. Proposed gels, with similar rheological properties, may be used for ex vivo and in vivo studies to obtain a suitable drug activity of metronidazole in the treatment of acne rosacea. [Abstract/Link to Full Text]

Bani-Jaber A, Hamdan I, Al-Khalidi B
Sodium mefenamate as a solution for the formulation and dissolution problems of mefenamic acid.
Chem Pharm Bull (Tokyo). 2007 Aug;55(8):1136-40.
Sodium salt formation of mefenamic acid (MA) was studied as a way to solve the formulation and dissolution problems of MA. For this purpose, sodium salt of mefenamic acid (Na-MA) was prepared by reacting MA powder with equimolar sodium hydroxide in an aqueous phase, and consequently, Na-MA solution was obtained. The resultant solution was lyophilized and Na-MA powder was collected. The salt formation was confirmed by the results of fourier transformation-infrared (FTIR) spectroscopy and differential scanning calorimetry (DSC) studies on Na-MA powder in comparison to MA powder. Na-MA powder was assessed for direct compressibility, in comparison to MA powder, when formulated as a mixture with minimum amount of Avicel((R)) pH 101 and then compressed into tablets using a hydraulic tablet press. Na-MA tablets exhibited satisfactory hardness and friability, and did not show capping or lamination. On the other hand, some MA tablets showed capping or lamination upon compression and all the tested MA tablets for friability capped. Na-MA tablets were also studied for drug dissolution, in comparison to MA tablets, in water, a pH 7.4 phosphate buffer, and a pH 7.4 phosphate buffer after soaking in 0.1 m HCl. Under these different dissolution conditions, Na-MA tablets showed much higher dissolution rate and extent than MA tablets. The results of the study suggested that Na-MA can be considered as a solution form for the formulation and dissolution problems of MA. [Abstract/Link to Full Text]

Shih MH, Su YS, Wu CL
Syntheses of aromatic substituted hydrazino-thiazole derivatives to clarify structural characterization and antioxidant activity between 3-arylsydnonyl and aryl substituted hydrazino-thiazoles.
Chem Pharm Bull (Tokyo). 2007 Aug;55(8):1126-35.
This work clarifies the structural characterization and antioxidant activity between aromatic and 3-arylsydnonyl substituted hydrazino-thiazoles by further synthesizing a series of aromatic ring-substituted hydrazino-thiazole derivatives 8a-h and 9a-h. Hydrazino-thiazole derivatives 8a-h and 9a-h were obtained by reacting aromatic or heterocyclic aromatic aldehyde thiosemicarbazones 7a-h with cyclization reagents ethyl 2-chloroacetoacetate (2a) and 2-bromoacetophenone (2b), respectively. The ORTEP drawings of compounds 8g, 8h and 9f provide strong evidence of the structure of aromatic thiazole derivatives 8a-h and 9a-h. Undoubtedly, the structure of compounds 3e-h and 4e-h synthesized by the reaction of 3-aryl-4-formylsydnone thiosemicarbazones 1e-h with cyclization reagents 2a and 2b in the previous work should have the thiazole moiety, and not the thiazoline moiety. Both the new thiazole derivatives 8a-h and 9a-h and the 3-arylsydnonyl-substituted derivatives 3e-h and 4e-h were investigated to determine their antioxidant activity by two tests that have been highly documented-the direct scavenging effect on a stable free 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical and the inhibition of the 2,2-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) radical. Results of this study demonstrate that not only the thiazole ring and the aryl ring has the contribution to the antioxidant activities, the sydnone ring of 3-arylsydnonyl moiety also has its considerable contribution. [Abstract/Link to Full Text]

Wang L, Cui FD, Sunada H
Improvement of the dissolution rate of nitrendipine using a new pulse combustion drying method.
Chem Pharm Bull (Tokyo). 2007 Aug;55(8):1119-25.
Solid dispersions (SDs) of nitrendipine (NTD), a poorly water-soluble drug, were prepared with the Hypulcon pulse combustion dryer system, and the physicochemical properties of particles were investigated and compared with those of particles prepared with a spray dryer. The SD particles prepared with Hypulcon using Aerosil and Tween 80 as carriers showed improved properties over those prepared with a conventional spray dryer, such as smaller particle size, tighter particle size distribution, and no agglomeration. Powder X-ray diffraction and differential scanning calorimetry evaluation showed that the drug in the NTD-Aerosil SD prepared with 5% (v/v) Tween 80 solution was dispersed in an amorphous state. Fourier transformation IR spectroscopy indicated the presence of hydrogen bonds between NTD and Aerosil. Aerosil had greater ability to improve the dissolution of NTD than Sylysia and other polymers. The highest drug supersaturation concentration was maintained continuously during the dissolution test of the NTD-Aerosil SD prepared with 5% (v/v) Tween 80 solution using Hypulcon. The good hydrophilicity and dispersibility of Aerosil, solubilization of Tween 80, and actions of shock waves and ultrasonic waves might account for the amorphization of NTD and improved dissolution rate of SDs. Pulse combustion drying with low drying costs and high thermal efficiency is a promising method for the preparation of SD particles with improved properties without using organic solvent. [Abstract/Link to Full Text]

Arisawa M
Development of environmentally benign organometallic catalysis for drug discovery and its application.
Chem Pharm Bull (Tokyo). 2007 Aug;55(8):1099-118.
We have developed a novel organometallic catalysis and applied it to drug discovery. Two new catalysts were found, ruthenium hydride with a nitrogen-containing heterocyclic carbene (A) and an organopalladium catalyst supported on a sulfur-terminated semiconductor, gallium arsenide (001) (B). Both catalysts are environmentally benign, because A can yield indole derivatives with good atom economy, and B can catalyze the Mizoroki-Heck reaction more than 10 times with only trace amounts of leached palladium (ppb level). We also describe our synthetic study of nitrogen-containing heterocycles using ring-closing metathesis (RCM), such as chiral bicyclic lactams, azacycloundecenes, axially chiral macrolactams, 1,2-dihydroquinolines and indoles, including the development of silyl-enol ether ene metathesis, selective isomerization of terminal olefin, enamide metathesis and cycloisomerization and its application to the syntheis of 4 natural products, (-)-coniceine, (S)-pyrrolam A, angustureine, and fistulosin. [Abstract/Link to Full Text]

El-Beih AA, Kawabata T, Koimaru K, Ohta T, Tsukamoto S
Monodictyquinone A: a new antimicrobial anthraquinone from a sea urchin-derived fungus Monodictys sp.
Chem Pharm Bull (Tokyo). 2007 Jul;55(7):1097-8.
A new antimicrobial anthraquinone, 1,8-dihydroxy-2-methoxy-6-methylanthraquinone, monodictyquinone A (1), was isolated from a culture of a marine-derived fungus of the genus Monodictys which was isolated from the sea urchin, Anthocidaris crassispina, along with three known compounds, pachybasin (2), chrysophanol (3), and emodin (4). [Abstract/Link to Full Text]

Ono M, Sugita F, Shigematsu S, Takamura C, Yoshimitsu H, Miyashita H, Ikeda T, Nohara T
Three new steroid glycosides from the underground parts of Trillium kamtschaticum.
Chem Pharm Bull (Tokyo). 2007 Jul;55(7):1093-6.
Three new steroid glycosides named trikamsterosides C, D and E were isolated from the underground parts of Trillium kamtschaticum PALL. (Liliaceae) along with two known 18-norspirostanol glycosides trillenosides A and B. Their chemical structures were determined on the basis of spectroscopic data and chemical evidence. [Abstract/Link to Full Text]

Hou LH, Chen DL, Jian XX, Wang FP
Three new diterpenoid alkaloids from roots of Aconitum ouvrardianum HAND.-MAZZ.
Chem Pharm Bull (Tokyo). 2007 Jul;55(7):1090-2.
A new C(19)-diterpenoid alkaloid, ouvrardiantine (1) and two new C(20)-diterpenoid alkaloids, ouvrardiandines A (2) and B (3) were isolated from the root of Aconitum ouvrardianum HAND.-MAZZ. The structure of the new alkaloids was established on the basis of spectral data (1D- and 2D-NMR, HR-MS). [Abstract/Link to Full Text]

Li F, Li W, Fu H, Zhang Q, Koike K
Pancreatic lipase-inhibiting triterpenoid saponins from fruits of Acanthopanax senticosus.
Chem Pharm Bull (Tokyo). 2007 Jul;55(7):1087-9.
Sixteen triterpenoid saponins were isolated from the fruits of Acanthopanax senticosus, including a new compound, acanthopanaxoside E (1), which was established as 3-O-beta-D-glucuronopyranosyl echinocystic acid 28-O-beta-D-glucopyranoside on the basis of various spectroscopic analyses and chemical degradation. By using a pancreatic lipase-inhibiting assay system, the crude saponin fraction showed inhibitory activity on pancreatic lipase, which is a key enzyme in lipid digestion. Among the isolated compounds, silphioside F (2), copteroside B (3), hederagenin 3-O-beta-D-glucuronopyranoside 6'-O-methyl ester (4) and gypsogenin 3-O-beta-D-glucuronopyranoside (5) showed inhibitory activity toward pancreatic lipase with IC(50) values of 0.22, 0.25, 0.26 and 0.29 mM, respectively, and the free carboxylic acid groups in position 28 within their chemical structures were required for enhancement of pancreatic lipase inhibition. [Abstract/Link to Full Text]

Li D, Ikeda T, Nohara T, Liu J, Wen Y, Sakamoto T, Nonaka G
Cucurbitane glycosides from unripe fruits of Siraitia grosvenori.
Chem Pharm Bull (Tokyo). 2007 Jul;55(7):1082-6.
Studies on the constituents of the unripe fruits of Siraitia grosvenori led to the isolation of three new cucurbitane triterpene glycosides, 11-oxomogroside III (10), 11-dehydroxymogroside III (11), and 11-oxomogroside IV A (12). Their structures were determined on the basis of detailed analyses of 1D, 2D-NMR spectroscopic methods. All of the compounds isolated from the unripe fruits of S. grosvenori were tested for cytotoxic activities against tumor cells, HCT-116 and SMMC-7721. [Abstract/Link to Full Text]

Matsushita S, Yanai Y, Fusyuku A, Ikeda T, Ono M, Nohara T
Distinction of absolute configuration at C-22 of C-23-hydroxyspirostane and C-23-hydroxyspirosolane glycosides.
Chem Pharm Bull (Tokyo). 2007 Jul;55(7):1079-81.
It has been revealed that the absolute configurations at C-22 of 23-hydroxyspirostane and 23-hydroxyspirosolane could be unambiguouly judged by the (1)H- and (13)C-NMR spectroscopies. [Abstract/Link to Full Text]

Matsushita S, Yanai Y, Fusyuku A, Fujiwara Y, Ikeda T, Ono M, Han C, Ojika M, Nohara T
Efficient conversion of tomatidine into neuritogenic pregnane derivative.
Chem Pharm Bull (Tokyo). 2007 Jul;55(7):1077-8.
Moderate acetylation of tomatidine with anhydrous acetic acid and pyridine for 20 h at r.t., followed by pseudomerization in ice-water, gave a delta(20(22))-pseudo compound, which was then subjected to ozonolysis to provide a pregnane derivative in a total 54% yield showing neuritogenic and NGF-enhancing activities. [Abstract/Link to Full Text]

Qian ZM, Li HJ, Li P, Ren MT, Tang D
Simultaneous qualitation and quantification of thirteen bioactive compounds in Flos lonicerae by high-performance liquid chromatography with diode array detector and mass spectrometry.
Chem Pharm Bull (Tokyo). 2007 Jul;55(7):1073-6.
A high-performance liquid chromatography (HPLC) with diode array detector (DAD) and electrospray ionization mass spectrometry (ESI-MS) was established for the simultaneous determination of thirteen bioactive compounds in Flos Lonicerae. The optimal chromatographic conditions were obtained on a C(18) column (250x4.6 mm, 5.0 microm) with the column temperature at 30 degrees C. The mobile phase was composed of (A) acetic acid aqueous (0.4%, v/v) and (B) acetonitrile using a gradient elution, the flow rate was 1 ml/min. Detection wavelengths were set at 240 nm for iridoids (loganin, sweroside, secoxyloganin and centauroside), 330 nm for phenolic acids (chlorogenic acid, caffeic acid, 4,5-di-O-caffeoyl quinic acid and 3,4-di-O-caffeoyl quinic acid) and 360 nm for flavonoids (rutin, hyperoside, quercetin-3-O-glucoside, luteolin-7-O-glucoside and lonicerin). The identities of the peaks were accomplished by comparing retention times, UV and mass data with reference compounds under the same conditions. All calibration curves showed good linear regression (r(2)>0.9983) within test ranges. The developed method provided satisfactory precision and accuracy with overall intra-day and inter-day variations of 0.78--1.85% and 1.13--2.36%, respectively, and the overall recoveries of 91.3--104.2% for the thirteen compounds analyzed. The verified method was successfully applied to quantitative determination of the three types of bioactive compounds in ten commercial Flos Lonicerae samples from different markets in China. The analytical results demonstrated that the contents of the thirteen analytes were relatively variant. [Abstract/Link to Full Text]

Iqbal A, Siddiqui HL, Ashraf CM, Bukhari MH, Akram CM
Synthesis, spectroscopic and cytotoxic studies of biologically active new schiff bases derived from p-nitrobenzaldehyde.
Chem Pharm Bull (Tokyo). 2007 Jul;55(7):1070-2.
Thirteen new Schiff bases derived from p-nitrobenzaldehyde were synthesized by condensation with the appropriate amines. An unusual reduction of the p-nitrobenzaldehyde to the corresponding alcohol was also observed in one of the reactions. The structures of the compounds were identified using spectroscopic techniques. Cytotoxicity for the titled compounds was studied against Brine Shrimp, used as the test animal. [Abstract/Link to Full Text]

Iida K, Nakamura M, Hanamitsu H, Kajiwara M
Identification of tetrapyrrole compounds excreted by Rhodobacter sphaeroides and sources of the methyl hydrogens of bacteriochlorophyll a biosynthesized by R. sphaeroides, based on 13C-NMR spectral analysis of coproporphyrin III tetramethyl ester.
Chem Pharm Bull (Tokyo). 2007 Jul;55(7):1067-9.
Red-fluorescent tetrapyrrole compounds excreted by Rhodobacter sphaeroides into the culture broth were concluded to be coproporphyrinogen (Copro'gen) III and uroporphyrinogen (Uro'gen) I, based on the (13)C-NMR spectral identification of coproporphyrin (Copro) III tetramethyl ester and uroproporphyrin (Uro) I octamethyl ester. The sources of the methyl hydrogens of bacteriochlorophyll a were established by analysis of the (13)C-NMR spectra of (2)H,(13)C-Copro III tetramethyl ester chemically derived from (2)H,(13)C-Copro'gen III biosynthesized through the feeding of delta-amino[2-(13)C]levulinic acid (ALA) to R. sphaeroides in medium containing 50% (2)H(2)O. We confirmed the previous finding that one of the methyl hydrogens was derived from water in the medium during decarboxylation of four acetyl side chains of Uro'gen III to generate Copro'gen III. It was further shown that the other hydrogen atoms, previously reported to be derived from methylene hydrogens at C-2 of ALA, had been exchanged with hydrogen of water in the medium in the biosynthetic pathways leading from ALA to Copro'gen III. [Abstract/Link to Full Text]

Park YJ, Kim HJ, Lee SJ, Choi HY, Jin C, Lee YS
A new chromone, 11-hydroxy-sec-O-glucosylhamaudol from Ostericum koreanum.
Chem Pharm Bull (Tokyo). 2007 Jul;55(7):1065-6.
From the ethyl acetate fraction of the roots of Ostericum koreanum, a new chromone, 11-hydroxy-sec-O-glucosylhamaudol (1) along with the known compounds: four chromones, three coumarins, six phenolic compounds, and three quinic acids were isolated. These compounds were assessed for antioxidant activities in the DPPH radical and superoxide anion radical scavenging assay systems. Among isolates, 4-(2-hydroxy-vinyl)-benzene-1,2-diol (12) showed the most potent DPPH radical scavenging activity (IC(50)=4.80+/-0.62 mug/ml) and superoxide anion radical scavenging activity (IC(50)=11.05+/-0.83 microg/ml) in the xanthine/xanthine oxidase system. The antioxidant activities of 12 were comparable to those of quercetin and luteolin. [Abstract/Link to Full Text]

Choshi T, Uchida Y, Kubota Y, Nobuhiro J, Takeshita M, Hatano T, Hibino S
Lipase-catalyzed asymmetric synthesis of desprenyl-carquinostatin A and descycloavandulyl-lavanduquinocin.
Chem Pharm Bull (Tokyo). 2007 Jul;55(7):1060-4.
An asymmetric synthesis of the core carbazole structure, 6-desprenyl-carquinostatin 3 and 6-descycloavandulyl-lavanduquinocin 3, toward a total synthesis of carquinostatin A (1) and lavanduquinocin (2), has been established. Lipase QLM (Meito) catalyzed enantioselective acetylation of the racemic alcohol 6 gave the (-)-acetate 7 and the (+)-alcohol 6 with high enantioselectivity. The absolute stereochemistry of the (-)- and (+)-alcohol 6 have been determined to be R- and S-configurations, respectively, by the advanced Mosher method. In the same manner, the (-)-acetate 13 and the (+)-alcohol 12 have been obtained from the racemic alcohol 12. The (R)-(-)-acetate 13, derived from the (R)-(-)-acetate 7, was the same as the (-)-acetate 13, which has been determined to be (R)-configuration. Oxidation of the (R)-(-)-acetate 13 followed by hydrolysis afforded (R)-(-)-6-desprenyl-carquinostatin [and (R)-(-)-6-descycloavandulyl-lavanduquinocin] 3. In addition, oxidation of the (S)-(+)-alcohol 12 provided (S)-(+)-3, which is the enantiomer of 6-desprenyl-carquinostatin A (R)-(-)-3. [Abstract/Link to Full Text]

Usui S, Fujieda H, Suzuki T, Yoshida N, Nakagawa H, Ogura M, Makishima M, Miyata N
Synthesis and evaluation of 2-Nonylaminopyridine derivatives as PPAR ligands.
Chem Pharm Bull (Tokyo). 2007 Jul;55(7):1053-9.
To find novel PPAR ligands, we prepared several 3-{3 or 4-[2-(nonylpyridin-2-ylamino)ethoxy]phenyl}propanoic acid derivatives which were designed based on the structure of our previous PPARgamma ligand 1. In PPAR binding affinity assays, compound 4, which had an ethoxy group at the C-2 position of the propanoic acid of 1, showed selective binding affinity for PPARgamma. Compound 3, with an ethyl group at the C-2 position, was found to be a PPARalpha/gamma dual ligand. Compound 6, the meta isomer of 1, has been shown to be a PPARalpha ligand. The introduction of methyl (7) and ethyl (8) groups to the C-2 position of the propanoic acid of 6 further improved PPARalpha-binding potency. In cell-based transactivation assay, compounds 3 and 4 showed dual-agonist activity toward PPARalpha and PPARgamma. Compound 6 was found to be a triple agonist and compound 8 proved to be a selective PPARalpha agonist. In the human hypodermic preadipocyte differentiation test, it was demonstrated that the maximal activity of compounds 3 and 4 was higher than that of rosiglitazone. [Abstract/Link to Full Text]

Kisa F, Yamada K, Miyamoto T, Inagaki M, Higuchi R
Determination of the absolute configuration of sialic acids in gangliosides from the sea cucumber Cucumaria echinata.
Chem Pharm Bull (Tokyo). 2007 Jul;55(7):1051-2.
Enantiomeric pairs of sialic acid, D- and L-NeuAc (N-acetylneuraminic acid), were converted to D- and L-arabinose, respectively, by chemical degradation. Using this method, the absolute configuration of the sialic acid residues, NeuAc and NeuGc (N-glycolylneuraminic acid), in the gangliosides from the sea cucumber Cucumaria echinata was determined to be the D-form. Although naturally occurring sialic acids have been believed to be the D-form on the basis of biosynthetic evidence, this is the first report of the determination of the absolute configuration of the sialic acid residues in gangliosides using chemical methods. [Abstract/Link to Full Text]

Nozawa D, Okubo T, Ishii T, Chaki S, Okuyama S, Nakazato A
Synthesis of diphenylmethyl analogues and their affinity for the melanocortin-4 receptor and the serotonin transporter.
Chem Pharm Bull (Tokyo). 2007 Jul;55(7):1044-50.
While examining antagonists of the melanocortin-4 receptor (MC4 receptor), we found that compound 12b, containing a diphenylmethyl moiety, had a relatively high affinity for the MC4 receptor. When diphenylmethyl analogues were further examined, compounds 12c and 18 were also found to exhibit a high affinity for the MC4 receptor (IC(50)=46.7 nM and 33.2 nM, respectively). Furthermore, compound 12c was also found to show a high affinity for the serotonin transporter (IC(50)=10.7 nM). Here, we describe the synthesis and biological evaluation of various diphenylmethyl analogues in relation to their actions on the MC4 receptor and the serotonin transporter. [Abstract/Link to Full Text]

Araya I, Tsubuki T, Saito T, Numata M, Akita H
Synthesis of the metabolites of 4-(2-methyl-1H-imidazol-1-yl)-2,2-diphenylbutanamide (KRP-197/ONO-8025).
Chem Pharm Bull (Tokyo). 2007 Jul;55(7):1039-43.
We synthesized the six presumed metabolites (2--7) of 4-(2-methyl-1H-imidazol-1-yl)-2,2-diphenylbutanamide [KRP-197/ONO-8025, 1], a urinary incontinence therapeutic agent, in order to confirm the structures of the metabolites. Metabolite (2) was synthesized via glucuronidaion of compound (1) and methyl 2,3,4-tri-O-benzoyl-1-methanesulfonyl-alpha-D-glucopyranuronate. Metabolite (3) was synthesized via 3-(tert-butoxycarbonyl)-2-methyl-1,3-imidazolidine-4,5-dione. Metabolites (4--7) were synthesized via 4-amino-2-diphenylbutanamide, respectively. The structures of the metabolites (2--7) in humans were identified by means of synthesis of the authentic compounds. [Abstract/Link to Full Text]

Yoshikawa M, Sugimoto S, Nakamura S, Sakumae H, Matsuda H
Medicinal flowers. XVI. New dammarane-type triterpene tetraglycosides and gastroprotective principles from flower buds of Panax ginseng.
Chem Pharm Bull (Tokyo). 2007 Jul;55(7):1034-8.
The oligoglycoside fraction from the flower buds of Panax ginseng C. A. MEYER (Araliaceae) was found to show protective effects on ethanol-induced gastric mucosal lesions in rats. From the oligoglycoside fraction, new dammarane-type triterpene tetraglycosides, floralginsenosides M, N, O, and P, were isolated together with the major oligoglycosides ginsenoside Rd and Re. The structures of the new floralginsenosides were elucidated on the basis of chemical and physicochemical evidence. Ginsenoside Rd (protopanaxadiol 3,20-O-bisdesmoside) exhibited inhibitory effects on ethanol- and indomethacin-induced gastric mucosal lesions in rats. [Abstract/Link to Full Text]

Hikosou R, Kurabayashi Y, Doumoto M, Hoshitoku K, Mizushima F, Minoura K, Tomoo K, Ishida T
Effect of DNA on filament formation of tau microtubule-binding domain: structural dependence of DNA.
Chem Pharm Bull (Tokyo). 2007 Jul;55(7):1030-3.
To examine whether or not DNA accelerates the paired helical filament (PHF) formation of tau, the effect of various types of DNAs on filament formations of three-repeated and four-repeated microtubule-binding domains (3RMBD and 4RMBD, respectively) of tau protein was investigated by monitoring the change of thioflavin S fluorescence intensity, that is parallel to the filament formation. Consequently, the followings were clarified: 1) the structurally rigid double-stranded DNA such as poly(dG-dC) or calf thymus DNA has the high potency of promoting the filament formations of 3RMBD and 4RMBD, 2) the filament formation of 3RMBD was more promoted than that of 4RMBD, due to the intermolecular dimer formation of 3RMBD, 3) the DNA-promoted filament formations of these MBDs were temperature-dependent, and the single-stranded DNA such as poly(dA) or poly(dT) reversely protected 4RMBD from the molecular assembly at 20 degrees C. These are the first report on the function of DNA for the PHF formation of tau protein. [Abstract/Link to Full Text]

Recent Articles in The Journal of Experimental Medicine

Gross S, Tilly P, Hentsch D, Vonesch JL, Fabre JE
Vascular wall-produced prostaglandin E2 exacerbates arterial thrombosis and atherothrombosis through platelet EP3 receptors.
J Exp Med. 2007 Feb 19;204(2):311-20.
Prostanoids, bioactive lipids derived from arachidonic acid (AA), are important for vascular homeostasis. Among them, prostaglandin E2 (PGE2) enhances aggregation of platelets submaximally stimulated in vitro. This results from activation of EP3, one of the four PGE2 receptors, which decreases the threshold at which agonists activate platelets to aggregate. Although PGE2 altered venous thrombosis induced by administration of AA, its role in pathophysiopathological conditions has remained speculative. We report that arterial walls subjected to inflammatory stimuli produce PGE2. In several models, we show that PGE2 produced by the arterial wall facilitates arterial thrombosis. Next, we detected PGE2 in mouse atherosclerotic plaques. We demonstrate that this plaque-produced PGE2 is not altered and is still able to activate EP3. In addition, we present evidence that PGE2 can leave the plaque and activate EP3 on blood platelets. Consistent with these findings, we observed that atherothrombosis induced in vivo by mechanical rupture of the plaque was drastically decreased when platelets lacked EP3. In conclusion, PGE2 facilitates the initiation of arterial thrombosis and, hence, contributes to atherothrombosis. Inhibition of the platelet EP3 receptor should improve prevention of atherothrombosis. [Abstract/Link to Full Text]

Bashyam H
IFNgamma: issuing macrophages a license to kill.
J Exp Med. 2007 Jan 22;204(1):3.
T cells tell macrophages when to start making the toxic soup of lysosomal enzymes, reactive oxygen species, and nitric oxide that destroys intracellular pathogens. In 1983, Carl Nathan proved that this start signal comes in the form of the secreted cytokine IFNgamma. [Abstract/Link to Full Text]

Hirota K, Hashimoto M, Yoshitomi H, Tanaka S, Nomura T, Yamaguchi T, Iwakura Y, Sakaguchi N, Sakaguchi S
T cell self-reactivity forms a cytokine milieu for spontaneous development of IL-17+ Th cells that cause autoimmune arthritis.
J Exp Med. 2007 Jan 22;204(1):41-7.
This report shows that highly self-reactive T cells produced in mice as a result of genetically altered thymic T cell selection spontaneously differentiate into interleukin (IL)-17-secreting CD4+ helper T (Th) cells (Th17 cells), which mediate an autoimmune arthritis that clinically and immunologically resembles rheumatoid arthritis (RA). The thymus-produced self-reactive T cells, which become activated in the periphery via recognition of major histocompatibility complex/self-peptide complexes, stimulate antigen-presenting cells (APCs) to secrete IL-6. APC-derived IL-6, together with T cell-derived IL-6, drives naive self-reactive T cells to differentiate into arthritogenic Th17 cells. Deficiency of either IL-17 or IL-6 completely inhibits arthritis development, whereas interferon (IFN)-gamma deficiency exacerbates it. The generation, differentiation, and persistence of arthritogenic Th17 cells per se are, however, insufficient for producing overt autoimmune arthritis. Yet overt disease is precipitated by further expansion and activation of autoimmune Th17 cells, for example, via IFN-gamma deficiency, homeostatic proliferation, or stimulation of innate immunity by microbial products. Thus, a genetically determined T cell self-reactivity forms a cytokine milieu that facilitates preferential differentiation of self-reactive T cells into Th17 cells. Extrinsic or intrinsic stimuli further expand these cells, thereby triggering autoimmune disease. Intervention in these events at cellular and molecular levels is useful to treat and prevent autoimmune disease, in particular RA. [Abstract/Link to Full Text]

Wang F, Langley R, Gulten G, Dover LG, Besra GS, Jacobs WR, Sacchettini JC
Mechanism of thioamide drug action against tuberculosis and leprosy.
J Exp Med. 2007 Jan 22;204(1):73-8.
Thioamide drugs, ethionamide (ETH) and prothionamide (PTH), are clinically effective in the treatment of Mycobacterium tuberculosis, M. leprae, and M. avium complex infections. Although generally considered second-line drugs for tuberculosis, their use has increased considerably as the number of multidrug resistant and extensively drug resistant tuberculosis cases continues to rise. Despite the widespread use of thioamide drugs to treat tuberculosis and leprosy, their precise mechanisms of action remain unknown. Using a cell-based activation method, we now have definitive evidence that both thioamides form covalent adducts with nicotinamide adenine dinucleotide (NAD) and that these adducts are tight-binding inhibitors of M. tuberculosis and M. leprae InhA. The crystal structures of the inhibited M. leprae and M. tuberculosis InhA complexes provide the molecular details of target-drug interactions. The purified ETH-NAD and PTH-NAD adducts both showed nanomolar Kis against M. tuberculosis and M. leprae InhA. Knowledge of the precise structures and mechanisms of action of these drugs provides insights into designing new drugs that can overcome drug resistance. [Abstract/Link to Full Text]

Ronai D, Iglesias-Ussel MD, Fan M, Li Z, Martin A, Scharff MD
Detection of chromatin-associated single-stranded DNA in regions targeted for somatic hypermutation.
J Exp Med. 2007 Jan 22;204(1):181-90.
After encounter with antigen, the antibody repertoire is shaped by somatic hypermutation (SHM), which leads to an increase in the affinity of antibodies for the antigen, and class-switch recombination (CSR), which results in a change in the effector function of antibodies. Both SHM and CSR are initiated by activation-induced cytidine deaminase (AID), which deaminates deoxycytidine to deoxyuridine in single-stranded DNA (ssDNA). The precise mechanism responsible for the formation of ssDNA in V regions undergoing SHM has yet to be experimentally established. In this study, we searched for ssDNA in mutating V regions in which DNA-protein complexes were preserved in the context of chromatin in human B cell lines and in primary mouse B cells. We found that V regions that undergo SHM were enriched in short patches of ssDNA, rather than R loops, on both the coding and noncoding strands. Detection of these patches depended on the presence of DNA-associated proteins and required active transcription. Consistent with this, we found that both DNA strands in the V region were transcribed. We conclude that regions of DNA that are targets of SHM assemble protein-DNA complexes in which ssDNA is exposed, making it accessible to AID. [Abstract/Link to Full Text]

Nimmerjahn F, Ravetch JV
The antiinflammatory activity of IgG: the intravenous IgG paradox.
J Exp Med. 2007 Jan 22;204(1):11-5.
How high doses of intravenous IgG (IVIG) suppress autoimmune diseases remains unresolved. We have recently shown that the antiinflammatory activity of IVIG can be attributed to a minor species of IgGs that is modified with terminal sialic acids on their Fc-linked glycans. Here we propose that these Fc-sialylated IgGs engage a unique receptor on macrophages that, in turn, leads to the upregulation of an inhibitory Fcgamma receptor (FcgammaR), thereby protecting against autoantibody-mediated pathology. [Abstract/Link to Full Text]

Liu J, Guan X, Ma X
Regulation of IL-27 p28 gene expression in macrophages through MyD88- and interferon-gamma-mediated pathways.
J Exp Med. 2007 Jan 22;204(1):141-52.
Interleukin (IL)-27 is the newest member of the IL-12 family of heterodimeric cytokines composed of the Epstein-Barr virus-induced gene 3 and p28 chains. IL-27 not only plays an important role in the regulation of differentiation of naive T helper cells but also possesses antiinflammatory properties. IL-27 is an early product of activated monocytes/macrophages and dendritic cells. However, the mechanisms whereby inflammatory signals stimulate IL-27 production have not been explored. In this study, we investigated the transcriptional regulation of the mouse IL-27 p28 gene in macrophages in response to lipopolysaccharide (LPS) and interferon (IFN)-gamma. We found that LPS-stimulated p28 production was completely dependent on the Toll-like receptor 4/myeloid differentiation factor 88 (MyD88)-mediated pathway but only partially dependent on nuclear factor kappaB c-Rel. IFN-gamma-induced p28 production/secretion was also partially dependent on MyD88 but independent of c-Rel. We then cloned the mouse p28 gene promoter and mapped its multiple transcription initiation sites. Furthermore, we identified critical promoter elements that mediate the inductive effects of LPS and IFN-gamma, separately and synergistically, on p28 gene transcription in a c-Rel- and interferon regulatory factor 1-dependent manner, respectively. [Abstract/Link to Full Text]

Villarino AV, Tato CM, Stumhofer JS, Yao Z, Cui YK, Hennighausen L, O'Shea JJ, Hunter CA
Helper T cell IL-2 production is limited by negative feedback and STAT-dependent cytokine signals.
J Exp Med. 2007 Jan 22;204(1):65-71.
Although required for many fundamental immune processes, ranging from self-tolerance to pathogen immunity, interleukin (IL)-2 production is transient, and the mechanisms underlying this brevity remain unclear. These studies reveal that helper T cell IL-2 production is limited by a classic negative feedback loop that functions autonomously or in collaboration with other common gamma chain (IL-4 and IL-7) and IL-6/IL-12 family cytokines (IL-12 and IL-27). Consistent with this model for cytokine-dependent regulation, they also demonstrate that the inhibitory effect can be mediated by several signal transducer and activator of transcription (STAT) family transcription factors, namely STAT5, STAT4, and STAT6. Collectively, these findings establish that IL-2 production is limited by a network of autocrine and paracrine signals that are readily available during acute inflammatory responses and, thus, provide a cellular and molecular basis for its transient pattern of expression. [Abstract/Link to Full Text]

Serafini M, Dylla SJ, Oki M, Heremans Y, Tolar J, Jiang Y, Buckley SM, Pelacho B, Burns TC, Frommer S, Rossi DJ, Bryder D, Panoskaltsis-Mortari A, O'Shaughnessy MJ, Nelson-Holte M, Fine GC, Weissman IL, Blazar BR, Verfaillie CM
Hematopoietic reconstitution by multipotent adult progenitor cells: precursors to long-term hematopoietic stem cells.
J Exp Med. 2007 Jan 22;204(1):129-39.
For decades, in vitro expansion of transplantable hematopoietic stem cells (HSCs) has been an elusive goal. Here, we demonstrate that multipotent adult progenitor cells (MAPCs), isolated from green fluorescent protein (GFP)-transgenic mice and expanded in vitro for >40-80 population doublings, are capable of multilineage hematopoietic engraftment of immunodeficient mice. Among MAPC-derived GFP+CD45.2+ cells in the bone marrow of engrafted mice, HSCs were present that could radioprotect and reconstitute multilineage hematopoiesis in secondary and tertiary recipients, as well as myeloid and lymphoid hematopoietic progenitor subsets and functional GFP+ MAPC-derived lymphocytes that were functional. Although hematopoietic contribution by MAPCs was comparable to control KTLS HSCs, approximately 10(3)-fold more MAPCs were required for efficient engraftment. Because GFP+ host-derived CD45.1+ cells were not observed, fusion is not likely to account for the generation of HSCs by MAPCs. [Abstract/Link to Full Text]

Zhang B, Bowerman NA, Salama JK, Schmidt H, Spiotto MT, Schietinger A, Yu P, Fu YX, Weichselbaum RR, Rowley DA, Kranz DM, Schreiber H
Induced sensitization of tumor stroma leads to eradication of established cancer by T cells.
J Exp Med. 2007 Jan 22;204(1):49-55.
Targeting cancer cells, as well as the nonmalignant stromal cells cross-presenting the tumor antigen (Ag), can lead to the complete destruction of well-established solid tumors by adoptively transferred Ag-specific cytotoxic T lymphocytes (CTLs). If, however, cancer cells express only low levels of the Ag, then stromal cells are not destroyed, and the tumor escapes as Ag loss variants. We show that treating well-established tumors expressing low levels of Ag with local irradiation or a chemotherapeutic drug causes sufficient release of Ag to sensitize stromal cells for destruction by CTLs. This was shown directly using high affinity T cell receptor tetramers for visualizing the transient appearance of tumor-specific peptide-MHC complexes on stromal cells. Maximum loading of tumor stroma with cancer Ag occurred 2 d after treatment and coincided with the optimal time for T cell transfer. Under these conditions, tumor rejection was complete. These findings may set the stage for developing rational clinical protocols for combining irradiation or chemotherapy with CTL therapy. [Abstract/Link to Full Text]

Casellas R, Zhang Q, Zheng NY, Mathias MD, Smith K, Wilson PC
Igkappa allelic inclusion is a consequence of receptor editing.
J Exp Med. 2007 Jan 22;204(1):153-60.
The discovery of lymphocytes bearing two light chains in mice carrying self-reactive antibody transgenes has challenged the "one lymphocyte-one antibody" rule. However, the extent and nature of allelically included cells in normal mice is unknown. We show that 10% of mature B cells coexpress both Igkappa alleles. These cells are not the result of failure in allelic exclusion per se, but arise through receptor editing. We find that under physiological conditions, editing occurs both by deletion and by inclusion with equal probability. In addition, we demonstrate that B lymphocytes carrying two B-cell receptors are recruited to germinal center reactions, and thus fully participate in humoral immune responses. Our data measure the scope of allelic inclusion and provide a mechanism whereby autoreactive B cells might "escape" central tolerance. [Abstract/Link to Full Text]

Tarbell KV, Petit L, Zuo X, Toy P, Luo X, Mqadmi A, Yang H, Suthanthiran M, Mojsov S, Steinman RM
Dendritic cell-expanded, islet-specific CD4+ CD25+ CD62L+ regulatory T cells restore normoglycemia in diabetic NOD mice.
J Exp Med. 2007 Jan 22;204(1):191-201.
Most treatments that prevent autoimmune diabetes in nonobese diabetic (NOD) mice require intervention at early pathogenic stages, when insulitis is first developing. We tested whether dendritic cell (DC)-expanded, islet antigen-specific CD4+ CD25+ suppressor T cells could treat diabetes at later stages of disease, when most of the insulin-producing islet beta cells had been destroyed by infiltrating lymphocytes. CD4+ CD25+ CD62L+ regulatory T cells (T reg cells) from BDC2.5 T cell receptor transgenic mice were expanded with antigen-pulsed DCs and IL-2, and were then injected into NOD mice. A single dose of as few as 5x10(4) of these islet-specific T reg cells blocked diabetes development in prediabetic 13-wk-old NOD mice. The T reg cells also induced long-lasting reversal of hyperglycemia in 50% of mice in which overt diabetes had developed. Successfully treated diabetic mice had similar responses to glucose challenge compared with nondiabetic NOD mice. The successfully treated mice retained diabetogenic T cells, but also had substantially increased Foxp3+ cells in draining pancreatic lymph nodes. However, these Foxp3+ cells were derived from the recipient mice and not the injected T reg cells, suggesting a role for endogenous T reg cells in maintaining tolerance after treatment. Therefore, inoculation of DC-expanded, antigen-specific suppressor T cells has considerable efficacy in ameliorating ongoing diabetes in NOD mice. [Abstract/Link to Full Text]

Lahl K, Loddenkemper C, Drouin C, Freyer J, Arnason J, Eberl G, Hamann A, Wagner H, Huehn J, Sparwasser T
Selective depletion of Foxp3+ regulatory T cells induces a scurfy-like disease.
J Exp Med. 2007 Jan 22;204(1):57-63.
The scurfy mutant mouse strain suffers from a fatal lymphoproliferative disease leading to early death within 3-4 wk of age. A frame-shift mutation of the forkhead box transcription factor Foxp3 has been identified as the molecular cause of this multiorgan autoimmune disease. Foxp3 is a central control element in the development and function of regulatory T cells (T reg cells), which are necessary for the maintenance of self-tolerance. However, it is unclear whether dysfunction or a lack of T reg cells is etiologically involved in scurfy pathogenesis and its human correlate, the IPEX syndrome. We describe the generation of bacterial artificial chromosome-transgenic mice termed "depletion of regulatory T cell" (DEREG) mice expressing a diphtheria toxin (DT) receptor-enhanced green fluorescent protein fusion protein under the control of the foxp3 gene locus, allowing selective and efficient depletion of Foxp3+ T reg cells by DT injection. Ablation of Foxp3+ T reg cells in newborn DEREG mice led to the development of scurfy-like symptoms with splenomegaly, lymphadenopathy, insulitis, and severe skin inflammation. Thus, these data provide experimental evidence that the absence of Foxp3+ T reg cells is indeed sufficient to induce a scurfy-like phenotype. Furthermore, DEREG mice will allow a more precise definition of the function of Foxp3+ T reg cells in immune reactions in vivo. [Abstract/Link to Full Text]

Kleinschek MA, Owyang AM, Joyce-Shaikh B, Langrish CL, Chen Y, Gorman DM, Blumenschein WM, McClanahan T, Brombacher F, Hurst SD, Kastelein RA, Cua DJ
IL-25 regulates Th17 function in autoimmune inflammation.
J Exp Med. 2007 Jan 22;204(1):161-70.
Interleukin (IL)-25 is a member of the IL-17 family of cytokines. However, unlike the other members of this family, IL-25 promotes T helper (Th) 2 responses. We now show that IL-25 also regulates the development of autoimmune inflammation mediated by IL-17-producing T cells. We have generated IL-25-deficient (il25-/-) mice and found that they are highly susceptible to experimental autoimmune encephalomyelitis (EAE). The accelerated disease in the il25-/- mice is associated with an increase of IL-23 in the periphery and a subsequent increase in the number of inflammatory IL-17-, IFNgamma-, and TNF-producing T cells that invade the central nervous system. Neutralization of IL-17 but not IFNgamma in il25-/- mice prevented EAE, suggesting that IL-17 is a major disease-promoting factor. IL-25 treatment at several time points during a relapse-remitting model or chronic model of EAE completely suppressed disease. IL-25 treatment induced elevated production of IL-13, which is required for suppression of Th17 responses by direct inhibition of IL-23, IL-1beta, and IL-6 expression in activated dendritic cells. Thus, IL-25 and IL-17, being members of the same cytokine family, play opposing roles in the pathogenesis of organ-specific autoimmunity. [Abstract/Link to Full Text]

Ito T, Yang M, Wang YH, Lande R, Gregorio J, Perng OA, Qin XF, Liu YJ, Gilliet M
Plasmacytoid dendritic cells prime IL-10-producing T regulatory cells by inducible costimulator ligand.
J Exp Med. 2007 Jan 22;204(1):105-15.
Although there is evidence for distinct roles of myeloid dendritic cells (DCs [mDCs]) and plasmacytoid pre-DCs (pDCs) in regulating T cell-mediated adaptive immunity, the concept of functional DC subsets has been questioned because of the lack of a molecular mechanism to explain these differences. In this study, we provide direct evidence that maturing mDCs and pDCs express different sets of molecules for T cell priming. Although both maturing mDCs and pDCs upregulate the expression of CD80 and CD86, only pDCs upregulate the expression of inducible costimulator ligand (ICOS-L) and maintain high expression levels upon differentiation into mature DCs. High ICOS-L expression endows maturing pDCs with the ability to induce the differentiation of naive CD4 T cells to produce interleukin-10 (IL-10) but not the T helper (Th)2 cytokines IL-4, -5, and -13. These IL-10-producing T cells are T regulatory cells, and their generation by ICOS-L is independent of pDC-driven Th1 and Th2 differentiation, although, in the later condition, some contribution from endogenous IL-4 cannot be completely ruled out. Thus, in contrast to mDCs, pDCs are poised to express ICOS-L upon maturation, which leads to the generation of IL-10-producing T regulatory cells. Our findings demonstrate that mDC and pDCs are intrinsically different in the expression of costimulatory molecules that drive distinct types of T cell responses. [Abstract/Link to Full Text]

Nadkarni S, Mauri C, Ehrenstein MR
Anti-TNF-alpha therapy induces a distinct regulatory T cell population in patients with rheumatoid arthritis via TGF-beta.
J Exp Med. 2007 Jan 22;204(1):33-9.
The induction of regulatory T (T reg) cells holds considerable potential as a treatment for autoimmune diseases. We have previously shown that CD4+CD25hi T reg cells isolated from patients with active rheumatoid arthritis (RA) have a defect in their ability to suppress proinflammatory cytokine production by CD4+CD25- [corrected] T cells. This defect, however, was overcome after anti-tumor necrosis factor (TNF)-alpha antibody (infliximab) therapy. Here, we demonstrate that infliximab therapy gives rise to a CD4+CD25hiFoxP3+ T reg cell population, which mediates suppression via transforming growth factor (TGF)-beta and interleukin 10, and lacks CD62L expression, thereby distinguishing this T reg cell subset from natural T reg cells present in healthy individuals and patients with active RA. In vitro, infliximab induced the differentiation of CD62L- T reg cells from CD4+CD25- T cells isolated from active RA patients, a process dependent on TGF-beta. In spite of the potent suppressor capacity displayed by this CD62L- T reg cell population, the natural CD62L+ T reg cells remained defective in infliximab-treated patients. These results suggest that anti-TNF-alpha therapy in RA patients generates a newly differentiated population of T reg cells, which compensates for the defective natural T reg cells. Therefore, manipulation of a proinflammatory environment could represent a therapeutic strategy for the induction of T reg cells and the restoration of tolerance. [Abstract/Link to Full Text]

Hua X, Kovarova M, Chason KD, Nguyen M, Koller BH, Tilley SL
Enhanced mast cell activation in mice deficient in the A2b adenosine receptor.
J Exp Med. 2007 Jan 22;204(1):117-28.
Antigen-mediated cross-linking of IgE bound to mast cells via the high affinity receptor for IgE triggers a signaling cascade that results in the release of intracellular calcium stores, followed by an influx of extracellular calcium. The collective increase in intracellular calcium is critical to the release of the granular contents of the mast cell, which include the mediators of acute anaphylaxis. We show that the sensitivity of the mast cell to antigen-mediated degranulation through this pathway can be dramatically influenced by the A2b adenosine receptor. Loss of this Gs-coupled receptor on mouse bone marrow-derived mast cells results in decreased basal levels of cyclic AMP and an excessive influx of extracellular calcium through store-operated calcium channels following antigen activation. Mice lacking the A2b receptor display increased sensitivity to IgE-mediated anaphylaxis. Collectively, these findings show that the A2b adenosine receptor functions as a critical regulator of signaling pathways within the mast cell, which act in concert to limit the magnitude of mast cell responsiveness when antigen is encountered. [Abstract/Link to Full Text]

Neuberger MS, Rada C
Somatic hypermutation: activation-induced deaminase for C/G followed by polymerase eta for A/T.
J Exp Med. 2007 Jan 22;204(1):7-10.
Somatic hypermutation (SHM) introduces nucleotide substitutions into immunoglobulin variable (Ig V) region genes at all four bases, but the mutations at C/G and A/T pairs are achieved by distinct mechanisms. Mutations at C/G pairs are a direct consequence of the C-->U deamination catalyzed by activation-induced deaminase (AID). Mutations at A/T pairs, however, require a second mutagenic process that occurs during patch repair of the AID-generated U/G mismatch. Several DNA polymerases have been proposed to play a role in SHM, but accumulating evidence indicates that the mutations at A/T are overwhelmingly achieved by recruitment of DNA polymerase eta. [Abstract/Link to Full Text]

Delbos F, Aoufouchi S, Faili A, Weill JC, Reynaud CA
DNA polymerase eta is the sole contributor of A/T modifications during immunoglobulin gene hypermutation in the mouse.
J Exp Med. 2007 Jan 22;204(1):17-23.
Mutations at A/T bases within immunoglobulin genes have been shown to be generated by a repair pathway involving the DNA-binding moiety of the mismatch repair complex constituted by the MSH2-MSH6 proteins, together with DNA polymerase eta (pol eta). However, residual A/T mutagenesis is still observed upon inactivation in the mouse of each of these factors, suggesting that the panel of activities involved might be more complex. We reported previously (Delbos, F., A. De Smet, A. Faili, S. Aoufouchi, J.-C. Weill, and C.-A. Reynaud. 2005. J. Exp. Med. 201:1191-1196) that residual A/T mutagenesis in pol eta-deficient mice was likely contributed by another enzyme not normally involved in hypermutation, DNA polymerase kappa, which is mobilized in the absence of the normal polymerase partner. We report the complete absence of A/T mutations in MSH2-pol eta double-deficient mice, thus indicating that the residual A/T mutagenesis in MSH2-deficient mice is contributed by pol eta, now recruited by uracil N-glycosylase, the second DNA repair pathway involved in hypermutation. We propose that this particular recruitment of pol eta corresponds to a profound modification of the function of uracil glycosylase in the absence of the mismatch repair complex, suggesting that MSH2-MSH6 actively prevent uracil glycosylase from error-free repair during hypermutation. pol eta thus appears to be the sole contributor of A/T mutations in the normal physiological context. [Abstract/Link to Full Text]

Riou C, Yassine-Diab B, Van grevenynghe J, Somogyi R, Greller LD, Gagnon D, Gimmig S, Wilkinson P, Shi Y, Cameron MJ, Campos-Gonzalez R, Balderas RS, Kelvin D, Sekaly RP, Haddad EK
Convergence of TCR and cytokine signaling leads to FOXO3a phosphorylation and drives the survival of CD4+ central memory T cells.
J Exp Med. 2007 Jan 22;204(1):79-91.
The molecular events involved in the establishment and maintenance of CD4+ central memory and effector memory T cells (TCM and TEM, respectively) are poorly understood. In this study, we demonstrate that ex vivo isolated TCM are more resistant to both spontaneous and Fas-induced apoptosis than TEM and have an increased capacity to proliferate and persist in vitro. Using global gene expression profiling, single cell proteomics, and functional assays, we show that the survival of CD4+ TCM depends, at least in part, on the activation and phosphorylation of signal transducer and activator of transcription 5a (STAT5a) and forkhead box O3a (FOXO3a). TCM showed a significant increase in the levels of phosphorylation of STAT5a compared with TEM in response to both IL-2 (P<0.04) and IL-7 (P<0.002); the latter is well known for its capacity to enhance T cell survival. Moreover, ex vivo TCM express higher levels of the transcriptionally inactive phosphorylated forms of FOXO3a and concomitantly lower levels of the proapoptotic FOXO3a target, Bim. Experiments aimed at blocking FOXO3a phosphorylation confirmed the role of this phosphoprotein in protecting TCM from apoptosis. Our results provide, for the first time in humans, an insight into molecular mechanisms that could be responsible for the longevity and persistence of CD4+ TCM. [Abstract/Link to Full Text]

Liu Y, Zhu M, Nishida K, Hirano T, Zhang W
An essential role for RasGRP1 in mast cell function and IgE-mediated allergic response.
J Exp Med. 2007 Jan 22;204(1):93-103.
Cross-linking of the FcepsilonRI activates the phosphatidyl inositol 3 kinase (PI3K) and mitogen-activated protein kinase pathways. Previous studies demonstrate that Ras guanyl nucleotide-releasing protein (RasGRP)1 is essential in T cell receptor-mediated Ras-Erk activation. Here, we report that RasGRP1 plays an important role in FcepsilonRI-mediated PI3K activation and mast cell function. RasGRP1-deficient mice failed to mount anaphylactic allergic reactions. RasGRP1-/- mast cells had markedly reduced degranulation and cytokine production. Although FcepsilonRI-mediated Erk activation was normal, PI3K activation was diminished. Consequently, activation of Akt, PIP3-dependent kinase, and protein kinase C delta was defective. Expression of a constitutively active form of N-Ras could rescue the degranulation defect and Akt activation. We further demonstrated that RasGRP1-/- mast cells were defective in granule translocation, microtubule formation, and RhoA activation. Our results identified RasGRP1 as an essential regulator of mast cell function. [Abstract/Link to Full Text]

Pua HH, Dzhagalov I, Chuck M, Mizushima N, He YW
A critical role for the autophagy gene Atg5 in T cell survival and proliferation.
J Exp Med. 2007 Jan 22;204(1):25-31.
Macroautophagy (hereafter referred to as autophagy) is a well-conserved intracellular degradation process. Recent studies examining cells lacking the autophagy genes Atg5 and Atg7 have demonstrated that autophagy plays essential roles in cell survival during starvation, in innate cell clearance of microbial pathogens, and in neural cell maintenance. However, the role of autophagy in T lymphocyte development and survival is not known. Here, we demonstrate that autophagosomes form in primary mouse T lymphocytes. By generating Atg5-/- chimeric mice, we found that Atg5-deficient T lymphocytes underwent full maturation. However, the numbers of total thymocytes and peripheral T and B lymphocytes were reduced in Atg5 chimeras. In the periphery, Atg5-/- CD8+ T lymphocytes displayed dramatically increased cell death. Furthermore, Atg5-/- CD4+ and CD8+ T cells failed to undergo efficient proliferation after TCR stimulation. These results demonstrate a critical role for Atg5 in multiple aspects of lymphocyte development and function and suggest that autophagy may be essential for both T lymphocyte survival and proliferation. [Abstract/Link to Full Text]

Varol C, Landsman L, Fogg DK, Greenshtein L, Gildor B, Margalit R, Kalchenko V, Geissmann F, Jung S
Monocytes give rise to mucosal, but not splenic, conventional dendritic cells.
J Exp Med. 2007 Jan 22;204(1):171-80.
The mononuclear phagocyte (MP) system is a body-wide macrophage (MPhi) and dendritic cell (DC) network, which contributes to tissue homeostasis, inflammation, and immune defense. The in vivo origins of MPs remain poorly understood. Here, we use an adoptive precursor cell transfer strategy into MP-depleted mice to establish the in vivo differentiation sequence from a recently identified MPhi/DC-restricted bone marrow (BM) precursor (MDP) via BM and blood intermediates to peripheral MPhis and DCs. We show that MDPs are in vivo precursors of BM and blood monocytes. Interestingly, grafted Gr1high "inflammatory" blood monocytes shuttle back to the BM in the absence of inflammation, convert into Gr1low monocytes, and contribute further to MP generation. The grafted monocytes give rise to DCs in the intestinal lamina propria and lung, but not to conventional CD11chigh DCs in the spleen, which develop during homeostasis from MDPs without a monocytic intermediate. [Abstract/Link to Full Text]

Harada M, Magara-Koyanagi K, Watarai H, Nagata Y, Ishii Y, Kojo S, Horiguchi S, Okamoto Y, Nakayama T, Suzuki N, Yeh WC, Akira S, Kitamura H, Ohara O, Seino K, Taniguchi M
IL-21-induced Bepsilon cell apoptosis mediated by natural killer T cells suppresses IgE responses.
J Exp Med. 2006 Dec 25;203(13):2929-37.
Epidemiological studies have suggested that the recent increase in the incidence and severity of immunoglobulin (Ig)E-mediated allergic disorders is inversely correlated with Mycobacterium bovis bacillus Calmette Guerin (BCG) vaccination; however, the underlying mechanisms remain uncertain. Here, we demonstrate that natural killer T (NKT) cells in mice and humans play a crucial role in the BCG-induced suppression of IgE responses. BCG-activated murine Valpha14 NKT cells, but not conventional CD4 T cells, selectively express high levels of interleukin (IL)-21, which preferentially induces apoptosis in Bepsilon cells. Signaling from the IL-21 receptor increases the formation of a complex between Bcl-2 and the proapoptotic molecule Bcl-2-modifying factor, resulting in Bepsilon cell apoptosis. Similarly, BCG vaccination induces IL-21 expression by human peripheral blood mononuclear cells (PBMCs) in a partially NKT cell-dependent fashion. BCG-activated PBMCs significantly reduce IgE production by human B cells. These findings provide new insight into the therapeutic effect of BCG in allergic diseases. [Abstract/Link to Full Text]

Wang Q, McLoughlin RM, Cobb BA, Charrel-Dennis M, Zaleski KJ, Golenbock D, Tzianabos AO, Kasper DL
A bacterial carbohydrate links innate and adaptive responses through Toll-like receptor 2.
J Exp Med. 2006 Dec 25;203(13):2853-63.
Commensalism is critical to a healthy Th1/Th2 cell balance. Polysaccharide A (PSA), which is produced by the intestinal commensal Bacteroides fragilis, activates CD4+ T cells, resulting in a Th1 response correcting the Th2 cell skew of germ-free mice. We identify Toll-like receptors as crucial to the convergence of innate and adaptive responses stimulated by PSA. Optimization of the Th1 cytokine interferon-gamma in PSA-stimulated dendritic cell-CD4+ T cell co-cultures depends on both Toll-like receptor (TLR) 2 and antigen presentation. Synergy between the innate and adaptive responses was also shown when TLR2-/- mice exhibited impaired intraabdominal abscess formation in response to B. fragilis. Commensal bacteria, using molecules like PSA, potentially modulate the Th1/Th2 cell balance and the response to infection by coordinating both the innate and adaptive pathways. [Abstract/Link to Full Text]

Yang SY, Fugmann SD, Schatz DG
Control of gene conversion and somatic hypermutation by immunoglobulin promoter and enhancer sequences.
J Exp Med. 2006 Dec 25;203(13):2919-28.
It is thought that gene conversion (GCV) and somatic hypermutation (SHM) of immunoglobulin (Ig) genes occur in two steps: the generation of uracils in DNA by activation-induced cytidine deaminase, followed by their subsequent repair by various DNA repair pathways to generate sequence-diversified products. It is not known how either of the two steps is targeted specifically to Ig loci. Because of the tight link between transcription and SHM, we have investigated the role of endogenous Ig light chain (IgL) transcriptional control elements in GCV/SHM in the chicken B cell line DT40. Promoter substitution experiments led to identification of a strong RNA polymerase II promoter incapable of supporting efficient GCV/SHM. This surprising finding indicates that high levels of transcription are not sufficient for robust GCV/SHM in Ig loci. Deletion of the IgL enhancer in a context in which high-level transcription was not compromised showed that the enhancer is not necessary for GCV/SHM. Our results indicate that cis-acting elements are important for Ig gene diversification, and we propose that targeting specificity is achieved through the combined action of several Ig locus elements that include the promoter. [Abstract/Link to Full Text]

Erlacher M, Labi V, Manzl C, Böck G, Tzankov A, Häcker G, Michalak E, Strasser A, Villunger A
Puma cooperates with Bim, the rate-limiting BH3-only protein in cell death during lymphocyte development, in apoptosis induction.
J Exp Med. 2006 Dec 25;203(13):2939-51.
The physiological role of B cell lymphoma 2 (Bcl-2) homology 3-only proteins has been investigated in mice lacking the individual genes identifying rate-limiting roles for Bim (Bcl-2-interacting mediator of cell death) and Puma (p53-up-regulated modulator of apoptosis) in apoptosis induction. The loss of Bim protects lymphocytes from apoptosis induced by cytokine deprivation and deregulated Ca++ flux and interferes with the deletion of autoreactive lymphocytes and the shutdown of immune responses. In contrast, Puma is considered the key mediator of p53-induced apoptosis. To investigate the hypothesis that Bim and Puma have overlapping functions, we generated mice lacking both genes and found that bim-/-/puma-/- animals develop multiple postnatal defects that are not observed in the single knockout mice. Most strikingly, hyperplasia of lymphatic organs is comparable with that observed in mice overexpressing Bcl-2 in all hemopoietic cells exceeding the hyperplasia observed in bim-/- mice. Bim and Puma also have clearly overlapping functions in p53-dependent and -independent apoptosis. Their combined loss promotes spontaneous tumorigenesis, causing the malignancies observed in Bcl-2 transgenic mice, but does not exacerbate the autoimmunity observed in the absence of Bim. [Abstract/Link to Full Text]

Wang XM, Zhang Y, Kim HP, Zhou Z, Feghali-Bostwick CA, Liu F, Ifedigbo E, Xu X, Oury TD, Kaminski N, Choi AM
Caveolin-1: a critical regulator of lung fibrosis in idiopathic pulmonary fibrosis.
J Exp Med. 2006 Dec 25;203(13):2895-906.
Idiopathic pulmonary fibrosis (IPF) is a progressive chronic disorder characterized by activation of fibroblasts and overproduction of extracellular matrix (ECM). Caveolin-1 (cav-1), a principal component of caveolae, has been implicated in the regulation of numerous signaling pathways and biological processes. We observed marked reduction of cav-1 expression in lung tissues and in primary pulmonary fibroblasts from IPF patients compared with controls. We also demonstrated that cav-1 markedly ameliorated bleomycin (BLM)-induced pulmonary fibrosis, as indicated by histological analysis, hydroxyproline content, and immunoblot analysis. Additionally, transforming growth factor beta1 (TGF-beta1), the well-known profibrotic cytokine, decreased cav-1 expression in human pulmonary fibroblasts. cav-1 was able to suppress TGF-beta1-induced ECM production in cultured fibroblasts through the regulation of the c-Jun N-terminal kinase (JNK) pathway. Interestingly, highly activated JNK was detected in IPF- and BLM-instilled lung tissue samples, which was dramatically suppressed by ad-cav-1 infection. Moreover, JNK1-null fibroblasts showed reduced smad signaling cascades, mimicking the effects of cav-1. This study indicates a pivotal role for cav-1 in ECM regulation and suggests a novel therapeutic target for patients with pulmonary fibrosis. [Abstract/Link to Full Text]

Ińiguez M, Berasain C, Martinez-Ansó E, Bustos M, Fortes P, Pennica D, Avila MA, Prieto J
Cardiotrophin-1 defends the liver against ischemia-reperfusion injury and mediates the protective effect of ischemic preconditioning.
J Exp Med. 2006 Dec 25;203(13):2809-15.
Ischemia-reperfusion (I/R) liver injury occurs when blood flow is restored after prolonged ischemia. A short interruption of blood flow (ischemic preconditioning [IP]) induces tolerance to subsequent prolonged ischemia through ill-defined mechanisms. Cardiotrophin (CT)-1, a cytokine of the interleukin-6 family, exerts hepatoprotective effects and activates key survival pathways like JAK/STAT3. Here we show that administration of CT-1 to rats or mice protects against I/R liver injury and that CT-1-deficient mice are exceedingly sensitive to this type of damage. IP markedly reduced transaminase levels and abrogated caspase-3 and c-Jun-NH2-terminal kinase activation after I/R in normal mice but not in CT-1-null mice. Moreover, the protective effect afforded by IP was reduced by previous administration of neutralizing anti-CT-1 antibody. Prominent STAT3 phosphorylation in liver tissue was observed after IP plus I/R in normal mice but not in CT-1-null mice. Oxidative stress, a process involved in IP-induced hepatoprotection, was found to stimulate CT-1 release from isolated hepatocytes. Interestingly, brief ischemia followed by short reperfusion caused mild serum transaminase elevation and strong STAT3 activation in normal and IL-6-deficient mice, but failed to activate STAT3 and provoked marked hypertransaminasemia in CT-1-null animals. In conclusion, CT-1 is an essential endogenous defense of the liver against I/R and is a key mediator of the protective effect induced by IP. [Abstract/Link to Full Text]

Shreeram S, Hee WK, Demidov ON, Kek C, Yamaguchi H, Fornace AJ, Anderson CW, Appella E, Bulavin DV
Regulation of ATM/p53-dependent suppression of myc-induced lymphomas by Wip1 phosphatase.
J Exp Med. 2006 Dec 25;203(13):2793-9.
The ataxia telangiectasia mutated (ATM) kinase is a key tumor suppressor that regulates numerous cell cycle checkpoints as well as apoptosis. Here, we report that ATM is a critical player in the regulation of apoptosis and lymphomagenesis in the presence of c-myc. In turn, deletion of the inhibitory ATM phosphatase, Wip1, results in ATM up-regulation and suppression of Emicro-myc-induced B cell lymphomas. Using mouse genetic crosses, we show that the onset of myc-induced lymphomas is dramatically delayed in Wip1-null mice in an ATM- and p53-, but not p38 MAPK- or Arf-, dependent manner. We propose that Wip1 phosphatase is critical for regulating the ATM-mediated tumor surveillance network. [Abstract/Link to Full Text]

Xu X, Zhang D, Zhang H, Wolters PJ, Killeen NP, Sullivan BM, Locksley RM, Lowell CA, Caughey GH
Neutrophil histamine contributes to inflammation in mycoplasma pneumonia.
J Exp Med. 2006 Dec 25;203(13):2907-17.
Mycoplasmas cause chronic inflammation and are implicated in asthma. Mast cells defend against mycoplasma infection and worsen allergic inflammation, which is mediated partly by histamine. To address the hypothesis that mycoplasma provokes histamine release, we exposed mice to Mycoplasma pulmonis, comparing responses in wild-type and mast cell-deficient KitW-sh/KitW-sh (W-sh) mice. Low histamine levels in uninfected W-sh mice confirmed the conventional wisdom that mast cells are principal sources of airway and serum histamine. Although mycoplasma did not release histamine acutely in wild-type airways, levels rose up to 50-fold above baseline 1 week after infection in mice heavily burdened with neutrophils. Surprisingly, histamine levels also rose profoundly in infected W-sh lungs, increasing in parallel with neutrophils and declining with neutrophil depletion. Furthermore, neutrophils from infected airway were highly enriched in histamine compared with naive neutrophils. In vitro, mycoplasma directly stimulated histamine production by naive neutrophils and strongly upregulated mRNA encoding histidine decarboxylase, the rate-limiting enzyme in histamine synthesis. In vivo, treatment with antihistamines pyrilamine or cimetidine decreased lung weight and severity of pneumonia and tracheobronchitis in infected W-sh mice. These findings suggest that neutrophils, provoked by mycoplasma, greatly expand their capacity to synthesize histamine, thereby contributing to lung and airway inflammation. [Abstract/Link to Full Text]

Liu X, Zhu Y, Dai S, White J, Peyerl F, Kappler JW, Marrack P
Bcl-xl does not have to bind Bax to protect T cells from death.
J Exp Med. 2006 Dec 25;203(13):2953-61.
Activated T cells die when antigen disappears from animals. This death is caused by proteins related to Bcl-2. Two hypotheses have been suggested to explain the actions of the different types of Bcl-2 proteins. One hypothesis suggests that, when T cells prepare to die, Bak and Bax, the proteins that actually kill activated T cells, are released from antiapoptotic proteins such as Bcl-2 and Bcl-xl. Another hypothesis suggests that Bak and Bax are normally free and are triggered to kill cells by release of messenger proteins, such as Bim, from Bcl-2 and Bcl-xl. Here, a form of Bcl-xl, which lacks a long unstructured loop, is used to show that the first hypothesis is not correct. Bcl-xl without its loop protects activated T cells from death, yet Bcl-xl without its loop cannot bind any form of Bak and Bax. Thus, binding of Bcl-xl to Bak or Bax is not involved in T cell life or death. The loop of Bcl-xl is also somewhat involved in Bcl-xl's binding of Bim because Bcl-xl without its loop binds Bim less well than wild-type Bcl-xl. Moreover, the loop may have additional, as yet unknown, functions because it changes its shape when Bcl-xl binds Bim. [Abstract/Link to Full Text]